Fig. 6.

SFN represses iRHOM2–p63 pathway in TOC. a Quantification of DHE staining by flow cytometry in TOC keratinocytes treated with dimethyl sulfoxide (DMSO; the vehicle control) and sulforaphane (SFN) (10 µM) for 24 h. b Growth curves of TOC cells cultured with SFN (10 µM) or DMSO for 0, 24 and 48 h post treatment. Data are expressed as mean ± SEM. c Percentage of apoptotic cells was quantified by using flow cytometry analysis of annexin-V-positive populations detected after 24 h incubation with SFN (10 µM) or DMSO, in TOC cells. Bars represent mean values with SEM of four experiments. d Immunoblotting of lysates from TOC keratinocytes treated with SFN (10 µM) or DMSO for 24 h and analysed for ΔNp63, iRHOM2, SURVIVIN, K16, p53, phopho-p53 and CYGB expression. GAPDH is used as a loading control. e Immunofluorescence staining of K16 in TOC cells after 0, 6 and 24 h of SFN incubation. Scale bar: 20 µm. f ELISA for TGFα and IL-6R with the supernatants of TOC cells treated with SFN (10 µM) or DMSO for 24 h. All data are expressed as mean and SEM of three experiments. Statistical analysis was performed by Student’s two-tailed t-test (*p < 0.05 and **p < 0.01)