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

Fig. 6.

Fig. 6.

Epfn promotes HaCaT cell differentiation through regulating Notch1. (A) Expression of keratinocyte marker genes as revealed by qPCR in HaCaT cells transfected with either Mock or Epfn expression vectors. (B) Co-immunoprecipitation assay of Epfn and E2F1 in HaCaT cells transfected with E2F1 (HA tag) and Epfn (VSV tag) expression vectors. IP, immunoprecipitated; IB, immunoblotted. (C) E2F activity assay in HaCaT cells transfected with Mock and Epfn expression vectors. (D) RBP-Jk reporter activity in HaCaT cells transfected with either Mock or Epfn expression vectors. Quantitative data in A,C,D show the mean+s.e.m. (three independent experiments); **P<0.01. (E) ChIP analysis of the Notch1 promoter using the anti-VSV antibody in HaCaT cells transfected with either Mock or Epfn (VSV tag) expression vectors. Upper panel, in silico prediction of potential Sp transcription factor binding sites (Sp sites A and B) in the human Notch1 promoter. Lower panel, nuclear extracts from either VSV–Epfn- or Mock-transfected HaCaT cells were used for ChIP analysis with primer sets for Sp sites A and B. ab, antibody.