Figure 7.

5-Aza-CdR inhibits ccRCC cell proliferation partially by demethylation of KLF5. (a–d) ShControl (ShCon) or ShKLF5 were stably expressed in 786-O or A498 cells, then cells were treated with or without 5-Aza-CdR (1 μM for 786-O cells and 10 μM for A498 cells) for 4 days. Western blots for DNMT1 and KLF5 expression (a and c) and CCK8 assay for cell proliferation (b and d) of these cells were shown. Cell growth curves were separately compared between ShCon with and without 5-Aza-CdR treatment or between ShCon and ShKLF5 with and without 5-Aza-CdR treatment. The statistical significance was indicated by P-value. (e–f) 786-O (e) or A498 cells (f) expressing ShCon and ShKLF5 were treated with different concentrations of 5-Aza-CdR and IC50 were calculated by GraphPad Prism 6.0 software. All experiments were repeated at least three times with triplicate samples. All bar graphs are plotted as mean±S.D. *P<0.05; **P<0.01; ***P<0.001 by two-tailed Student’s t-test