Fig. 4.

Human keratinocytes silenced for HK2 phenocopy p63-depleted cells. (A) Real-time measurements (mean ± SEM; n = 3; *P < 0.01) of OCR were performed as in Fig. 2 after 48 h of p63 and HK2 silencing (Left). The relative quantification of OCR and ECAR evaluated as percentage of control (Scr) are displayed in histograms (Middle and Right). (B) MitoSOX Red and JC1 staining confirm the similarities between sip63 and siHK2 phenotypes. Silencing HK2 in Hekn resembles the increase of superoxide anion levels and mitochondrial membrane polarization observed in p63-depleted cells (mean ± SD; n = 3; *P < 0.01). (C) HK2-depleted keratinocytes display reduced proliferative potential. EdU staining by flow cytometry demonstrates a 50% reduction of proliferation 48 h after Hekn transfection (Left). A clonogenic assay that transfected human keratinocytes with the Scramble control and siHK2 confirms a reduction in colony formation after 12 d of culture. The micrographs shown are representative images of one of three independent experiments.