Figure 2. Prohibition of senescence and contact inhibition mediated by p16^INK4a by weekly knockdown of p120-Kaiso siRNAs from Day 7.

HCECs cultured in MESCM up to Day 49 were monitored by phase contrast and cytolocalization of p16^INK4a by immunostaining (A). Cell apoptosis was demonstrated by green fluorescence using CellEvent^TM Caspase-3/7 Green Detection Reagent with nuclear counterstaining with Hoechst 33342 on Day 49 (A, Scale bars: 25 μm). The expression of senescence markers was measured by qRT-PCR [n = 3, *p < 0.05 and **p < 0.01, compared to the control without treatment (B) or with scrambled RNA (scRNA) at day 21 (C)]. The percentage of senescent cells were determined by β-galactosidase staining (D).