SETD2 inactivation prevents PTECs from senescence by active E2F signaling. (A) GFP and β-gal staining results of NT-day 20, SETD2-day 20 and SETD2-day 40 PTEC cultures. Representative microscopic views are shown. Quantification of GFP- and β-gal–positive cells was performed by using ImageJ software (National Institutes of Health, Bethesda, MD). The results are present as mean ± SD value of three independent experiments (right panels). (B) The mRNA expression of CDKN2A and E2F1 in SETD2-WT PTECs at day 6 (WT-day 6), SETD2-WT PTECs at day 16 (WT-day 16), and SETD2-KD PTECs at day 25 (KD-day 25) was determined by RT-qPCR. The expression level of target genes was normalized to RP II. The results are presented as 2−∆Ct values of three independent experiments with mean ± SD. One-way ANOVA with Dunnett multiple testing corrections showed significant differences between WT-day 6 and KD-day 25 PTECs compared with nonproliferating WT-day 16 PTECs. *P < .05, **P < .01.