Human/Mouse/Rat Phospho-HSP27 (S78/S82) Antibody

Detection of HSP27 by Western Blot

Decrease in PP2A activity increases phosphorylation of p38MAPK, pMAPKAPK2 and Hsp27 in CD133+ cells.(A upper panel) Immunoblot analysis and (A lower panel) PP2A activity analysis for HT-29 CD133+ and CD133− cells after MACS separation at day 4 of exposure to hypoxia and serum depletion. (B upper panel) Immunoblot analysis and (B lower panel) PP2A activity of CCS cells cultured under serum depletion in the presence of EGF (10 ng/mL) and FGF2 (10 ng/mL) (EGF+FGF2) for indicated time periods. The fold values are relative to the band intensity of the 5 day treatment. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23185379), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of HSP27 by Western Blot

Decrease in PP2A activity increases phosphorylation of p38MAPK, pMAPKAPK2 and Hsp27 in CD133+ cells.(A upper panel) Immunoblot analysis and (A lower panel) PP2A activity analysis for HT-29 CD133+ and CD133− cells after MACS separation at day 4 of exposure to hypoxia and serum depletion. (B upper panel) Immunoblot analysis and (B lower panel) PP2A activity of CCS cells cultured under serum depletion in the presence of EGF (10 ng/mL) and FGF2 (10 ng/mL) (EGF+FGF2) for indicated time periods. The fold values are relative to the band intensity of the 5 day treatment. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23185379), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of HSP27 by Western Blot

The involvement of Hsp27 activation in the anti-apoptosis pathway of CD133+ cells.Cells were lentivirally transfected with Hsp27 shRNA (shR1 and shR2) or scrambled shRNA, then exposed to hypoxia and serum depletion. CD133+ and CD133− cells were isolated using MACS separation 4 days later. (A) Immunoblot analysis for HT-29 protein levels of CD133+ and CD133− cells after MACS separation. (B) The percentage of CD133+ cells were analyzed by flow cytometry and the increased CD133+ percentage compared to CD133+ percentage in normoxia and growth medium was calculated. (C) Cells were re-exposed to hypoxia and serum depletion for 1 day, followed by TUNEL staining. (D-F) HT-29 cells were exposed to serum depletion and hypoxia in the presence of Quercetin, KRIBB3, or DMSO (vehicle control) for 4 days. (D) CD133+ cell were isolated and the protein levels were analyzed by western blotting. (E) The increased percentage of CD133+ cells and (F) apoptosis of CD133+ cells were analyzed by flow cytometry and TUNEL staining, respectively. Error bars represent standard deviations. (**p<0.01 compared with the scrambled as determined by the Student’s t test.) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23185379), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of HSP27 by Western Blot

The involvement of Hsp27 activation in the anti-apoptosis pathway of CD133+ cells.Cells were lentivirally transfected with Hsp27 shRNA (shR1 and shR2) or scrambled shRNA, then exposed to hypoxia and serum depletion. CD133+ and CD133− cells were isolated using MACS separation 4 days later. (A) Immunoblot analysis for HT-29 protein levels of CD133+ and CD133− cells after MACS separation. (B) The percentage of CD133+ cells were analyzed by flow cytometry and the increased CD133+ percentage compared to CD133+ percentage in normoxia and growth medium was calculated. (C) Cells were re-exposed to hypoxia and serum depletion for 1 day, followed by TUNEL staining. (D-F) HT-29 cells were exposed to serum depletion and hypoxia in the presence of Quercetin, KRIBB3, or DMSO (vehicle control) for 4 days. (D) CD133+ cell were isolated and the protein levels were analyzed by western blotting. (E) The increased percentage of CD133+ cells and (F) apoptosis of CD133+ cells were analyzed by flow cytometry and TUNEL staining, respectively. Error bars represent standard deviations. (**p<0.01 compared with the scrambled as determined by the Student’s t test.) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23185379), licensed under a CC-BY license. Not internally tested by R&D Systems.