Anti-TWEAKR Rabbit Monoclonal Antibody

Th17/Treg cell differentiation ratio and TWEAK/Fn14 signaling level in AC mice. ( A ) The state of mice ocular surface was observed by slit-lamp. ( B ) The clinical scores of mice were performed according to the status of eyelid, conjunctiva and cornea. ( C ) HE staining was utilized to evaluate the pathological changes of conjunctival tissue in mice. ( D ) The proportion of Th17 or Treg cells in spleen of mice was assessed by flow cytometry. ( E ) The levels of Th17 and Treg cytokines in mice spleen were evaluated by ELISA. ( F-H ) TWEAK and Fn14 levels in ocular conjunctival tissue were measured by qRT-PCR and WB. ( I ) IHC was used to observe the protein level of TWEAK and Fn14 in ocular conjunctival tissue. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Control
Index in PubMed under a CC BY license. PMID: 39592944

TWEAK knockdown affected conjunctivitis in AC mice. ( A ) The effect of TWEAK knockdown on the state of mice ocular surface was observed under slit-lamp. ( B ) Clinical score of eyelid, conjunctiva and cornea to assess the effect of TWEAK knockdown in AC mice. ( C-D ) HE staining and TB staining were utilized to evaluate the effect of TWEAK knockdown on conjunctivitis in mice. ( E-G ) The levels of TWEAK and Fn14 in conjunctival tissue were detected by qRT-PCR, IHC and WB. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Con + AAV-shNC; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. AC + AAV-shNC; ns, no significant difference
Index in PubMed under a CC BY license. PMID: 39592944

TWEAK regulated the Th17/Treg cell ratio in AC mice. ( A ) The effect of TWEAK knockdown on Th17 and Treg cell ratio in spleen of AC mice was observed by flow cytometry. ( B-C ) WB and IHC were employed to evaluate the expression levels of FoxP3 and RORγt in mice conjunctival tissue. ( D ) The effect of TWEAK knockdown on the levels of Th17 and Treg cytokines in mice spleen were evaluated by ELISA. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Con + AAV-shNC; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. AC + AAV-shNC; ns, no significant difference
Index in PubMed under a CC BY license. PMID: 39592944

TWEAK knockdown promoted Nrf2/HO-1 signaling pathway in AC mice. ( A ) qRT-PCR was utilized to measure the effect of TWEAK knockdown on Nrf2 and HO-1 mRNA levels in conjunctival tissue of mice. ( B-C ) The effect of TWEAK knockdown on protein levels of Nrf2 and HO-1 in conjunctival tissue of mice were detected by WB and IHC. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Con + AAV-shNC; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. AC + AAV-shNC; ns, no significant difference
Index in PubMed under a CC BY license. PMID: 39592944

Inhibition of Nrf2/HO-1 signaling pathway affected the improvement of conjunctivitis in AC mice from TWEAK knockdown. ( A ) The effect of Nrf2 inhibitor on ocular surface status was observed by slit-lamp in AC mice with TWEAK knockdown. ( B ) Clinical score of eyelid, conjunctiva and cornea to assess the effect of Nrf2 inhibitor in AC mice with TWEAK knockdown. ( C-D ) HE staining and TB staining were employed to evaluate the effect of Nrf2 inhibitor on conjunctivitis in AC mice with TWEAK knockdown. ( E-G ) The levels of Nrf2 and HO-1 in conjunctival tissue were detected by qRT-PCR, IHC and WB. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. AC + AAV-shNC; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. AC + AAV-shTWEAK
Index in PubMed under a CC BY license. PMID: 39592944

Inhibition of Nrf2/HO-1 signaling pathway affected Th17/Treg cell ratio in AC mice with TWEAK knockdown. ( A ) The effect of Nrf2 inhibitor on Th17/Treg cell ratio in AC mice with TWEAK knockdown was assessed by flow cytometry. ( B-C ) WB and IHC assays were employed to evaluate the expression of FoxP3 and RORγt in conjunctival tissue of AC mice. ( D ) The levels of Th17 and Treg cytokines in mice spleen were detected by ELISA. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. AC + AAV-shNC; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. AC + AAV-shTWEAK
Index in PubMed under a CC BY license. PMID: 39592944

Effects of ISO on the TWEAK/Fn14 signaling pathway in TGF-β1-induced HK-2 cells. (A, B) ISO directly interacts with Fn14, dose–response sensorgrams, and the affinity constant ( K d = 5.47 μM) of ISO with Fn14. (C–F) Western blot analysis of TWEAK, Fn14, and the phosphorylation levels of ERK1/2 in TGF-β1-induced HK-2 cells. β-actin was used as the loading control. Data are expressed as mean ± SD ( n = 3). # p < 0.05, ## p < 0.01 versus the sham group, * p < 0.05, ** p < 0.01 versus the UUO group.
Index in PubMed under a CC BY license. PMID: 40977693

Effect of ISO on the expression of TWEAK/Fn14 signaling pathway-related proteins in UUO rats. Representative images and quantification of Western blot results of the protein expression of TWEAK, Fn14, TRAF2, and BRAF (A–D) , and phosphorylation of MER1/2 and ERK1/2 (E–H) . β-actin was used as the loading control for total protein. (I–L) Relative optical density analysis of the nucleoprotein expression of p-SP1, Snail, and β-catenin. PCNA was used as the loading control for nucleoprotein. All data are expressed as the mean ± SD ( n = 5). ## p < 0.01 versus the sham group, * p < 0.05, ** p < 0.01 versus the UUO group.
Index in PubMed under a CC BY license. PMID: 40977693

Fn14 overexpression compromises the therapeutic effects of ISO and the regulation of ISO on the TWEAK/Fn14 pathway-related proteins on UUO model rats. (A, B) Serum Scr and BUN levels in different groups. (C, D) Representative images and the CVF of Masson staining (×400). (E–H) Representative and quantification analysis of Col III and FN immunohistochemical staining (×400). Nuclei were counterstained with DAPI (blue). (I–K) Western blot and the relative optical densities analysis of E-cadherin and α-SMA in kidney tissue. (L–O) Representative images and quantification of Western blot results of the protein expression of TWEAK, Fn14, and phosphorylation of ERK1/2. β-actin was used as the loading control. (P–R) Relative optical density analysis of the nucleoprotein expression of Snail and β-catenin. PCNA was used as the loading control for nucleoprotein. Data were shown as mean ± SD ( n = 3). ## p < 0.01 versus the sham group, * p < 0.05, ** p < 0.01 versus the UUO group, △ p < 0.05, △△ p < 0.01 versus the ISO group.
Index in PubMed under a CC BY license. PMID: 40977693

Western blot analysis of TWEAKR using anti-TWEAKR antibody (M06261).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human HUVEC whole cell lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: rat PC-12 whole cell lysates,
Lane 4: rat C6 whole cell lysates,
Lane 5: mouse NIH/3T3 whole cell lysates,
Lane 6: mouse 4T1 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-TWEAKR antigen affinity purified monoclonal antibody (Catalog # M06261) at 1:1000 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 at a dilution of 1:500 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for TWEAKR at approximately 17,20 kDa. The expected band size for TWEAKR is at 17,20 kDa.