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. 2014 Dec 15;127(24):5261–5272. doi: 10.1242/jcs.156778

Fig. 4.

Fig. 4.

Epfn expression in differentiated HaCaT cells and the role of Epfn in EGF-mediated cell proliferation. (A) Immunoblot analysis of Epfn and filaggrin expression in differentiating HaCaT cells cultured in a low-Ca2+ medium (KGM) with the addition of 0 mM, 0.7 mM or 1.2 mM Ca2+ for 4 days (4d) or 7 days (7d). (B) HaCaT cell proliferation. (a) Upper panel shows western blot analysis of endogenous Epfn expression in HaCaT cells transfected with control shRNA and Epfn shRNA. Lower panel shows densitometric analysis of the western blot assays using NIH ImageJ. (b) Proliferation of HaCaT cells transfected with control shRNA and Epfn shRNA. (C) EGF-mediated proliferation of HaCaT cells transfected with control shRNA and Epfn shRNA. (D) EGF-mediated proliferation of human epidermal keratinocytes transfected with control shRNA and Epfn shRNA. (E) EGF-mediated proliferation of keratinocytes from the wild-type (Epfn+/+) and Epfn−/− epidermis. Primary keratinocytes were cultured in KGM for 4 days with or without recombinant EGF (10 ng/ml). Quantitative data show the mean+s.e.m. (three independent experiments); *P<0.05; **P<0.01.