Anti-Phospho-AKT1 (S129) Rabbit Monoclonal Antibody

All lanes use the Antibody at 1:1K dilution for 1 hour at room temperature.

All lanes use the Antibody at 1:1K dilution for 1 hour at room temperature.

Western blot analysis of Phospho-AKT1 (S129) expression in (1) MCF-7 cell lysate treated with Alkaline Phosphatase; (2) MCF-7 cell lysate.

PTEN/PI3K/Akt pathway’s contribution in miR-21 induced proliferation in c-kit + CSCs. Cultured c-kit + CSCs were treated with miR-21 mimics for 48 h before subjected to EdU immunofluorescence (A–B), flow cytometry (C) or Western blot (D). To test the contribution of PTEN/PI3K/Akt signaling, PTEN and PI3K were inhibited with Phen or LY294002 respectively. (A) c-kit + CSCs were double stained by EdU (green) and DAPI (blue), and observed under a fluorescence microscope (Olympus). Bar = 50 µm. DAPI = propidium iodide. (B) The statistics of EdU positive CSCs from immunofluorescence in (A). n = 6 in each group. (C) Flow cytometry was employed to detect cell cycle profiles in CSCs underwent different treatments miR-21 mimics or Phen increased the proportion of S phase CSCs compared with Control or scramble treated groups. n = 3. (D) PTEN/PI3K/Akt pathway’s influences on BrdU expression, which was detected with immune blotting. Just like miR-21 mimics’ effect on BrdU, when PTEN was inhibited by Phen, there was notably increase of BrdU compared with Normal or Scramble group. When PI3K was inhibited by LY294002, there was notably decrease of BrdU in mimics+LY294002 group compared with mimics group in CSCs. n = 3 in each group. a, P < 0.05 compared with Control; b, P < 0.05 compared with Scramble; c, P < 0.05 compared with miR-21 mimics group; d, P < 0.05 compared with Phen group. Download full-size image DOI:
Index in PubMed under a CC BY license. PMID: 28168101

Expression change of PTEN/PI3K/Akt pathway in the process of miR-21 mimics induced proliferation in c-kit + CSCs. Cultured CSCs were treated with miR-21 mimics for 48 h before the subsequent procedures. To test the contribution of PTEN/PI3K/Akt signaling to miR-21 mimics’s pro-proliferation effects in c-kit + CSCs, PTEN and PI3K were inhibited with Phen or LY294002 respectively. (A) RT-PCR was carried out to detect miR-21 mimics’s effects on PTEN expression at the mRNA level, which showed no change between Control, miR-21 scramble, miR-21 mimics and miR-21 mimics+ LY294002 group, while Phen resulted in a significant down-regulation of PTEN compared with the other groups. (B–C) Western blot was carried out to detect miR-21 mimics’s effects on PTEN protein expression, which showed that miR-21 mimics significantly down-regulated PTEN protein in miR-21 mimics group compared with the scramble group. In addition, both Phen treatment and miR-21 mimics incubation increased p-Akt level, while PI3K inhibitor LY294002 decreased p-Akt level dramatically ( P < 0.05). a, P < 0.05 compared with Control; b, P < 0.05 compared with miR-21 scramble group; c, P < 0.05 compared with miR-21 mimics group; d, P < 0.05 compared with Phen group. n = 3 in each group. p-Akt = phosphor-Akt. Download full-size image DOI:
Index in PubMed under a CC BY license. PMID: 28168101

Shh and AKT signaling pathway inhibitors abolish the effect of EGCG on the growth of DPCs and ORSCs. (A,B) After treatment with EGCG and/or cyclopamine, GANT61 or LY294002, the cell viability of DPCs and ORSCs was assessed by MTT assay. (C,D) Protein levels of cyclinB1 and cyclinD1 in DPCs and ORSCs were assessed by western blot with β-actin as the internal reference. All experiments were repeated three times. The results are presented as mean ± SD. ### p < 0.001 compared with the control group; ∗∗∗ p < 0.001 compared with the EGCG group.
Index in PubMed under a CC BY license. PMID: 29997505

Epigallocatechin-3-gallate activates the Shh and AKT signaling pathways in DPCS and ORSCs. Upon treatment with EGCG, the protein levels of Shh (A) , PTCH (B) , Smo (C) , and Gli1 (D) in DPCs and ORSCs were detected by western blot with β-actin as the internal reference. (E) Western blot was performed to assess the levels of AKT and p-AKT in each group with β-actin as the internal reference. (F) Chemical structure of EGCG. Each experiment was repeated three times.
Index in PubMed under a CC BY license. PMID: 29997505

Epigallocatechin-3-gallate activates the Shh and AKT signaling pathways in hair follicles. After treatment with 0.5 and 1 μM EGCG, the mRNA levels of Shh (A) , PTCH (B) , Smo (C) , and Gli1 (D) in hair follicles were detected by qRT-PCR. The relative mRNA levels were calculated using 2 -ΔΔC t method. Western blot was also performed to detect the protein levels of Shh (E) , PTCH (F) , Smo (G) , and Gli1 (H) in hair follicles. β-actin served as the internal reference. (I) Level of Shh in hair follicles was assessed by immunofluorescence. Red fluorescence: Shh; blue fluorescence: DAPI. (J) The phosphorylation level of AKT was assessed by western blot. (K) Level of p-AKT in hair follicles was assessed by immunofluorescence. Red fluorescence: p-AKT; blue fluorescence: DAPI. Each experiment was repeated three times and the results are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 compared with the control group.
Index in PubMed under a CC BY license. PMID: 29997505

Effects of magnolol on mice alcohol-induced liver damage in the AKT/PI3K signaling pathway. Liver tissues were extracted for protein analysis by western blotting. AKT and PI3K, proteins expression were detected. The levels of AKT and PI3K were compared with GAPDH. The data were demonstrated as means ± SD. (*P < 0.05, **P < 0.01, ***P < 0.001).
Index in PubMed under a CC BY license. PMID: 31920652