Research Paper Volume 14, Issue 20 pp 8179—8204

Figure 4. The SIX2/SIRT1/AKT/GSK3β network can be activated by resveratrol and regulates the cell fate of UdRPCs. Cell culture medium of UdRPCs was supplemented with different concentrations of resveratrol for 24h. Activation of SIRT1 and phosphorylation of H2A.X were monitored by immunofluorescence-based detection (A) (scale bars: 100 μm). mRNA expression of SIRT1, ATM, P16 and SIX2 was determined in U48 by quantitative real-time PCR (B). Cell growth of U48 was evaluated after 24h of resveratrol treatment as depicted (C). Relative protein expression normalized to ß-ACTIN for SIRT1, AKT and GSK3ß and relative protein phosphorylation for AKT, GSK3ß and pH2A.X in U48 was detected by Western blot (D). RT-PCR analysis reveal Progerin transcripts in U48 treated with high concentrations of resveratrol (E). mRNA expression of SIRT1, ATM, P16 and SIX2 was determined in U51 by quantitative real-time PCR (F). Cell growth of U51 was evaluated after 24h of resveratrol treatment as depicted (G). Relative protein expression normalized to ß-ACTIN for SIRT1, AKT and GSK3ß and relative protein phosphorylation for AKT, GSK3ß and pH2A.X in U51 was detected by Western blot (H). RT-PCR analysis reveal Progerin transcripts in U51 treated with high concentrations of resveratrol (I).