Figure 2.

Ectopic expression of FOXM1 inhibits the differentiation response to loss of p53. (a) Immunodetection of the epidermal differentiation markers in red, involucrin (Invol) or keratin K1 (K1) in keratinocytes 3 days after infection with CT1 or shp53-1 and CT2 or FOX (as in Figure 1). DAPI for DNA in blue. (b) Bar histogram shows cells with high size and complexity (light scatter; differentiation), or positive for Invol or K1 relative to control cells (CT1/CT2=100%) as measured by flow cytomety. Bottom histograms show representative plots for the expression of K1 (+, positive cells according to negative isotype antibody control, red broken line). More details in Supplementary Figures 1b and c. (c) Real-time (RT)–PCR for expression of the differentiation markers Filaggrin (Filag) and Invol 2 days after infections as indicated (fold differences relative to control CT1/CT2). (d) Shedding cells collected from the culture supernatant due to differentiation 5 days after infections relative to control (CT1/CT2=100%). (e) Detection by western blot of β1 integrin 5 days after infections as indicated. GAPDH (GAP) was used as loading control. Scale bar: 50 μm; *P<0.05 and **P<0.01. Data are mean±s.e.m. of triplicate samples of representative experiments (n=3). Results are representative of two different strains from different individuals (N=2).