Anti-Bax Antibody Picoband®

Western blot analysis of Bax using anti-Bax antibody (A00183).
Electrophoresis was performed on a 12% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human THP-1 whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human Hela whole cell lysates,
Lane 5: rat C6 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Bax antigen affinity purified polyclonal antibody (A00183) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for Bax at approximately 21 kDa. The expected band size for Bax is at 21 kDa.

IHC analysis of Bax using anti-Bax antibody (A00183).
Bax was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Bax Antibody (A00183) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Bax using anti-Bax antibody (A00183).
Bax was detected in a paraffin-embedded section of mouse kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Bax Antibody (A00183) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Bax using anti-Bax antibody (A00183).
Bax was detected in a paraffin-embedded section of rat kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Bax Antibody (A00183) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of Bax using anti-Bax antibody (A00183).
Bax was detected in an immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-Bax Antibody (A00183) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of BAX using anti-BAX antibody (A00183).
BAX was detected in a paraffin-embedded section of FFPE mouse uterus tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with rabbit anti-BAX Antibody (A00183) at 10 μg/ml overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of THP-1 cells using anti-Bax antibody (A00183).
Overlay histogram showing THP-1 cells stained with A00183 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Bax Antibody (A00183, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

a Western blotting was used to examine mitochondrial apoptotic pathway-related proteins after treatment with control (i), PANI-PEG-CS (ii), Ir(III) complex (iii), and Ir(III)@PANI-PEG-CS (iv). Key proteins involved in apoptosis such as Bax, Bcl-2, cytochrome c, and cleaved caspase-3 were examined to better understand how each treatment affects cell death at the mitochondrial level. b To further investigate the underlying molecular mechanisms, western blotting was used to investigate the PI3K/AKT/mTOR pathway (i), PANI-PEG-CS (ii), Ir(III) complex (iii), and Ir(III)@PANI-PEG-CS (iv). Expression levels of PI3K, AKT (total and phosphorylated), and mTOR were evaluated to assess whether this survival pathway was activated or suppressed. Protein levels were quantified and compared to the control group to determine statistical significance. * p < 0.05, ** p < 0.01, *** p < 0.001 in comparison to the respective control
Index in Springer Nature under a CC BY license. DOI: 10.1186/s12645-025-00336-z

( A, B ) Protein expression levels of p21, p53, Bax, and Bcl-2 were detected by Western blot assays and compared by quantitative analysis of the gray value. * p <0.05 and ** p <0.01 vs. siR-NC group. siR-NC – control siRNA; siR-HMGB3 – HMGB3 siRNA; HMGB3 – high mobility group-box 3.
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( A ) Expression of Bcl-2 (A1–A3), ( B ) Bax (B1–B3), ( C ) Caspase-3 (C1–C3) in ovaries of HEV inoculation rabbits. A1, B1 and C1 are control group. A2, B2, C2 are HEV RNA positive ovaries in 28 dpi. A3, B3, C3 are HEV RNA positive ovaries in 49 dpi. (Original magnification:40×).
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Celastrol attenuates ganglion cells apoptosis in the retina of EAE rats. Treatment of celastrol decreased the number of TUNEL-positive cells (A) , upregulated expression of Bcl-2 (B) and downregulated expression of Bax, cleaved-caspase 3 and cleaved-PARP. Scale bar: 100 μm. Data were shown as mean ± SD, n = 5. ∗∗ P < 0.01 versus control group, ## P < 0.01 versus EAE group, †† P < 0.01 versus low dosage of celastrol group.
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Effects of maltol on the levels of inflammation cytokines in cisplatin-induced renal toxicity. ( A ) Effects of maltol on the positive expressions of Bax, Bcl-2, iNOS and COX-2 in renal tissues were examined by IHC in renal tissues (magnification × 200), And the column chart shows stained area, semiquantitative analysis of Bax, Bcl-2, iNOS and COX-2 expression in kidneys to IHC. ( B ) Inflammation cytokines level of TNF-α, IL-1β, iNOS and NF-κB in serum of mice were measured by ELISA kits. All values were expressed as mean ± S.D. * p < 0.05, ** p < 0.01 vs . normal group; # p < 0.05, ## p < 0.01 vs . cisplatin group.
Index in PubMed under a CC BY license. PMID: 30374107

V protein overexpression in DF-1 cells inhibited apoptosis through the Bcl-2\Bax-Caspase-3 pathway. A V and control (pCAGEN-flag) plasmids were transfected into DF-1 cells for 48 h before the cells were harvested. The RNA was extracted according to the previously described method. The mRNA levels of some apoptosis-related genes (proapoptosis genes Caspase-3, Caspase-9, and FasLG, and the antiapoptosis gene Bcl-2) were detected using Q-PCR. B DF-1 cells were transfected with V and control (pCAGEN-flag) plasmids for 48 h. Whole-cell extracts were prepared for Western blot analysis that was specific for the indicated proteins.
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Anti-SEMA4D antibody can inhibit the survival of AML cell lines in vivo and in vitro. A CCK-8 analysis of U937 and Molm-13 cells treated with VX15/2503 or not. B Cell apoptosis rate of U937 and Molm-13 cells treated with VX15/2503 or not was detected by flow cytometry using Annexin V-APC/PI staining. C Western blotting analysis was used to determine the expression of apoptosis-related proteins (Bcl-2, Bax, and cleaved-caspase3) in U937 and Molm-13 cells treated with VX15/2503 or not. Results of densitometry analysis of relative expression levels after normalization to loading control β-actin are presented. D Western blotting analysis was used to determine the expression of p-PI3K, PI3K, p-Akt, Akt in U937 and Molm-13 cells treated with VX15/2503 or not. Results of densitometry analysis of relative expression levels after normalization to loading control β-actin are presented. E Schematic outline of the mouse model delineating this experiment. F Nude mice were subcutaneously inoculated with U937 cells to establish AML xenograft tumors and treated with VX15/2503. Volumes of tumors were monitored by direct measurement. G Tumor size of xenograft mice in two groups. H Weights of tumors of xenograft mice in two groups. I Immunohistochemistry stain was used to measure the expression of Bcl-2, Bax, cleaved-caspase3, p-PI3K, p-Akt in xenograft tumors. J Western blotting analysis was used to determine the expression of Bcl-2, Bax, cleaved-caspase3, p-PI3K, PI3K, p-Akt, Akt in xenograft tumors. Results of densitometry analysis of relative expression levels after normalization to loading control β-actin are presented. Data with statistical significance are as indicated, *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant
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(A) Proliferation ability of MDA-MB-231 and MCF-7 induced by TGF-β1 by plate cloning experiment. The effect of TGF-β1 on the proliferation of MDA-MB-231 cells (B) and MCF-7 (C) cells were analyzed by CCK-8 (*p < 0.05, **p < 0.01). Analysis of the effect of TGF-β1 on the apoptosis of MDA-MB-231 cells (D) and MCF-7 (E) cells by Annexin V-FITC/PI stain flow cytometry (*p < 0.05, **p < 0.01). The expression level of the apoptosis and TP63 proteins in MDR-MB-231 cells (F) and MCF-7 (G) cells with TGF-β1 induced. GAPDH was used as an internal control. Quantitative analysis of TP63, Bcl-2 and Bax are expressed as the mean ± SD. *, **p < 0.01 vs. control group.
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SEMA4D promotes proliferation and inhibits apoptosis of AML cells. A Western blot was used to detect SEMA4D protein level when U937 and Molm-13 cells were transfected with stably knocking down or overexpressing SEMA4D lentivirus. B CCK-8 analysis of U937 and Molm-13 cells transfected with lentivirus targeting SEMA4D or control. C Colony formation assay of U937 and Molm-13 cells transfected with lentivirus targeting SEMA4D or control. D Cell apoptosis rate of U937 and Molm-13 cells transfected with lentivirus targeting SEMA4D or control was detected by flow cytometry using Annexin V-APC/PI staining. E Western blotting analysis was used to determine the expression of apoptosis-related proteins (Bcl-2, Bax, and cleaved-caspase3) in U937 and Molm-13 cells transfected with lentivirus targeting SEMA4D or control. Results of densitometry analysis of relative expression levels after normalization to loading control β-actin are presented. Data with statistical significance are as indicated, *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant
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SEMA4D functions through its receptor PlexinB1. A Western blot was used to detect PlexinB1 protein level when U937 and Molm-13 cells were transfected with siRNA-PlexinB1 or siRNA-control. B CCK-8 analysis of U937 and Molm-13 cells transfected with lentivirus targeting SEMA4D when PlexinB1 was knocked down or not. C Western blotting analysis was used to determine the expression of apoptosis-related proteins (Bcl-2, Bax, and cleaved-caspase3) in U937 and Molm-13 cells transfected with lentivirus targeting SEMA4D when PlexinB1 was knocked down or not. Results of densitometry analysis of relative expression levels after normalization to loading control β-actin are presented. D Cell apoptosis rate of U937 and Molm-13 cells transfected with lentivirus targeting SEMA4D when PlexinB1 was knocked down or not was detected by flow cytometry using Annexin V-APC/PI staining. E Western blotting analysis was used to determine the expression of p-PI3K, PI3K, p-Akt, Akt in U937 and Molm-13 cells transfected with lentivirus targeting SEMA4D when PlexinB1 was knocked down or not. Results of densitometry analysis of relative expression levels after normalization to loading control β-actin are presented. Data with statistical significance are as indicated, *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant
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Key proteins of apoptosis are upregulated in ACLF rats. ( A ) Western blot analysis for the BAX, CASP3, C-CASP3, CASP7, C-CASP7, CASP8, C-CASP8, and GAPDH proteins. Normal group: Lanes 1 to 3, ACLF group: Lanes 4 to 6. Relative expression level of ( B ) CASP3/GAPDH ( C ) C-CASP3/GAPDH ( D ) CASP7/GAPDH ( E ) C-CASP7/GAPDH ( F ) BAX/GAPDH ( G ) CASP8/GAPDH ( H ) C-CASP8/GAPDH. * P < 0.05 and ** P < 0.01. n = 3 per group.
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CircHIPK3 promotes H/R-induced cardiomyocyte apoptosis. A The intracellular ROS level was detected by flow cytometry. n = 3. B Annexin V-FITC/PI flow cytometry was used to evaluate the effect of circHIPK3 on cardiomyocyte apoptosis. n = 3. C Apoptosis-related proteins, including procaspase-3, cleaved caspase-3, Bax, and Bcl-2, were detected by western blotting. n = 3. * P < 0.05 compared with the normal group. # P < 0.05 compared with the H/R group.
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MiR-20b-5p inhibits autophagy and apoptosis of cardiomyocytes under H/R conditions. A Transfection efficacy of miR-20b-5p mimics and miR-20b-5p inhibitors in cardiomyocytes. B Western blot showed the effect of transfection of miR-20b-5p mimics and miR-20b-5p inhibitors on the expression of LC3II and P62 in normal cardiomyocytes. n = 3. C Western blot showed the effect of transfection of miR-20b-5p mimics and miR-20b-5p inhibitors on the expression of apoptosis-related proteins, including procaspase-3, cleaved caspase-3, Bax, and Bcl-2 in normal cardiomyocytes. n = 3. D Western blot showed the effects of miR-20b-5p mimics and miR-20b-5p inhibitor transfection on the expression of LC3II and P62 in cardiomyocytes under H/R conditions. n = 3. E Annexin V-FITC/PI flow cytometry was used to evaluate the effect of miR-20b-5p mimics and miR-20b-5p inhibitors on cardiomyocyte apoptosis under H/R conditions. n = 3. F Western blot analyzed the expression of apoptosis-related proteins, including procaspase-3, cleaved caspase-3, Bax, and Bcl-2 in cardiomyocytes under H/R conditions by miR-20b-5p mimics and miR-20b-5p inhibitors transfection. n = 3. * P < 0.05 compared with the mimics-NC (MNC) group. # P < 0.05 compared with the inhibitors-NC (INC) group.
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Protective effects of maltol on cisplatin-induced injury in HEK293 cells. ( A ) The cytotoxic effects of cisplatin on HEK293 cells. ( B ) Effect of maltol on the activity of normal cells. ( C ) The viability of HEK293 cells incubated with maltol after cisplatin exposure. Effects of maltol on the protein expression levels of Bcl-2, Bax and caspase 3, 8, 9 as well as GAPDH protein was used as a loading control. ( D ) Cells were used for western blot analysis of indicated proteins (upper panel). Column chart represents relative protein levels compared with the control group after normalization to GAPDH levels (lower panel) Values are expressed as mean ± S.D. n = 8. ** p < 0.01 vs . normal group; # p < 0.05, ## p < 0.01 vs . cisplatin group.
Index in PubMed under a CC BY license. PMID: 30374107

CircHIPK3 regulates autophagy and apoptosis via the CircHIPK3/miR-20b-5p/ATG7 axis. A Luciferase reporter assay showed that miR-20b-5p mimics directly binds to the 3′-UTR of ATG7 and inhibits luciferase activity. * P < 0.05 compared with the NC-mimics group. B ATG7 protein expression was detected by western blotting. n = 3. * P < 0.05 compared with the MNC group. # P < 0.05 compared with the INC group. C The protein expression levels of LC3-II, P62, and ATG7 were measured by western blotting. n = 3. D The intracellular ROS level was detected by flow cytometry. n = 3. E Annexin V-FITC/PI flow cytometry was used to evaluate apoptosis under different treatment conditions. n = 3. F Apoptosis-related proteins, including procaspase-3, cleaved caspase-3, Bax, and Bcl-2, were detected by western blotting. n = 3. * P < 0.05 compared with the H/R group. # P < 0.05 compared with the H/R + sicircHIPK3 group.
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TXNL1 induces apoptosis in DF-1 cells through the Bcl-2\Bax-Caspase-3 pathway. A Dot plot showing the flow cytometric analysis of phosphatidylserine (PS) translocation after staining with annexin V and PI in mock, control (pCMV-HA), and TXNL1-transfected DF-1 cells at 48 h. A representative of three independent experiments is shown. The lower right quadrant represents the early apoptotic cells (annexin v-positive), while the upper right quadrant shows the late apoptotic and necrotic cell populations (annexin v and PI-positive). B Dot plot showing the flow cytometric analysis of PS translocation after staining with annexin V and PI in mock, NC, and si290-transfected DF-1 cells at 36 h. C pCMV-HA-TXNL1 and control plasmids were transfected into DF-1 cells for 48 h before the cells were harvested. The RNA was extracted according to the previously described method. The mRNA level of some apoptosis-related genes (proapoptosis genes Caspase-3, Caspase-9, and FasLG and the antiapoptosis gene Bcl-2) were detected using Q-PCR. D DF-1 cells were transfected with pCMV-HA-TXNL1 and control plasmids for 48 h. Whole-cell extracts were prepared for Western blot analysis that was specific for the proteins indicated. E TXNL1 and control were transfected into DF-1 cells for 48 h before the cells were harvested. The mRNA levels of some interferon-related genes (IFN-α, IFN-β, IFN-γ, IRF1, IRF3) were detected using Q-PCR. F Si290 and NC were transfected into DF-1 cells for 36 h before the cells were harvested. The mRNA level of some apoptosis-related genes were detected using Q-PCR. G DF-1 cells were transfected with si290 and NC for 36 h. Whole-cell extracts were prepared for Western blot analysis that was specific for the indicated proteins. H si290 and NC were transfected into DF-1 cells for 36 h before the cells were harvested. The mRNA levels of some interferon-related genes were detected using Q-PCR.
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The influences of circPOSTN silencing on proliferation, apoptosis and aerobic glycolysis of glioma cells. a – l LN229 and U251 cells were transfected with si-circPOSTN or si-NC. a The interference efficiency of si-circPOSTN was analyzed with RT-qPCR assay in LN229 and U251 cells. b , c Effect of circPOSTN silencing on the cell viability of LN229 and U251 cells was assessed with MTT assay. d The apoptosis rate was computed with flow cytometry assay in transfected LN229 and U251 cells. e The western blot assay showed the expression levels of Bcl-2 and Bax in LN229 and U251 cells. f The caspase-3 activity was measured with a caspase-3 assay kit. g – i The concentration of glucose and lactate in the culture medium, as well as ATP production level were measured with a series of kits, respectively. j The protein expression levels of HK2 and LDHA were determined with western blot assay in transfected LN229 and U251 cells. k – l LDHA enzyme activity and ROS accumulation were evaluated in LN229 and U251 cells post-transfection with lactate dehydrogenase activity detection kit and reactive oxygen species assay kit, respectively. * P < 0.05
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Expression level of protein of Bcl-2, Bax and Caspase-3 in ovary tissue of different groups at 28 dpi and 49 dpi. ( A ) intensity of expression of Bcl-2 ( B ) Bax, ( C ) Caspase-3 in ovaries of HEV inoculation rabbits. ( D ) Western blot analysis of Bcl-2, Bax and Caspase-3 in ovaries in different groups.
Index in PubMed under a CC BY license. PMID: 29435117

Knockdown of circPOSTN mediated-effects on proliferation and apoptosis of glioma cells could be eliminated by silencing miR-361-5p. a – j LN229 and U251 cells were transfected with si-NC, si-circPOSTN, si-circPOSTN + anti-miR-NC, or si-circPOSTN + anti-miR-361-5p. a , b The relativity expression level of miR-361-5p was analyzed with RT-qPCR assay in LN229 and U251 cells. c , d MTT assay was administrated to assess cell viability of LN229 and U251 cells after transfection. e , f The apoptosis of transfected LN229 and U251 cells was monitored by flow cytometry. g , h The western blot assay was employed to show the expression levels of Bcl-2 and Bax in LN229 and U251 cells. i , j The caspase-3 activity was examined by caspase-3 assay kit. * P < 0.05
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TGF-β1 suppresses the HCC cells proliferative capacity but does not promote apoptosis. SMMC-7721 and BEL-7402 cells were treated with 0, 5ng/mL and 10ng/mL TGF-β1 for 48 hours. A . apoptosis B . western blotting examined the expression levels of Bcl-2 and Bax. Data are expressed as the mean ± SEM of three independent experiments, #P>0.05.
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TPX2 regulated proliferation, apoptosis, and aerobic glycolysis in glioma cells. a – l LN229 and U251 cells were introduced with si-NC or si-TPX2. a The transfection efficiency of si-TPX2 was checked with RT-qPCR assay in LN229 and U251 cells. b , c The cell viability of LN229 and U251 cells was determined with MTT assay. d The apoptosis rate of transfected LN229 and U251 cells was represented by flow cytometry assay. e The western blot assay was used to assay the expression levels of Bcl-2 and Bax in LN229 and U251 cells. f The activity of caspase-3 was detected with a caspase-3 assay kit. g – i The glucose, lactate, and ATP production levels were shown. j The protein expression levels of HK2 and LDHA were estimated by western blot assay in LN229 and U251 cells. k , l LDHA enzyme activity and ROS content were evaluated in LN229 and U251 cells post-transfection. * P < 0.05
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The effect of the combination of alteronol and ADM on protein levels of apoptosis-related molecules in 4T1 cells. (A) The protein levels of Bax and Bcl-2 were measured by western blot. (B) Quantitative analysis of Bax and Bcl-2 protein levels in 4T1 cells after treatment with alteronol and/or ADM. (C) The protein levels of cleaved PARP, cleaved caspase-9, and cleaved caspase-3 were examined by western blot. (D) Quantitative analysis of cleaved PARP, cleaved caspase-9, and cleaved caspase-3 protein levels after the indicated treatments. ∗ P < 0.05, ∗∗ P < 0.01 vs. control group. # P < 0.05, ## P < 0.01 vs. alteronol group. & P < 0.05, && P < 0.01 vs. ADM group. All data are expressed as mean ± SD of three independent experiments.
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