Research Paper Volume 9, Issue 10 pp 2052—2068

PKCζ-dependent upregulation of p27kip1 contributes to oxidative stress induced retinal pigment epithelial cell multinucleation

Figure 6. Inhibition of p27kip1 using Alsterpaullone, 2-Cyanoethyl (A2CE) and its effect on RPE multinucleation. (A) representative western blot from oxPOS, control, 5μM A2CE and 1μM A2CE treated RPE cells showing p27kip1 at 28 KDa and β-actin at 40 KDa. (B) quantification of p27kip1 protein expression by RPE cells in the different treatment groups. Signals for POS treated group were set to one and the data are represented as relative fold change to the POS group. * P <0.05, *** P <0.001 compared to the POS group. One-way ANOVA followed by Tukey’s multiple comparison test. N= 3. C-F: confocal images of multinucleate RPE cells in different treatment groups (C); POS-treated, (D); 1μM A2CE, (E); untreated RPE cells, (F); 5μM A2CE). (G) histogram showing the percentage of binucleate/multinucleate RPE cells following different treatments, * P <0.05, *** P <0.001 compared to oxPOS treated group. Data comprised of percentage of multinucleate cells per well from 5 different well for each group.