Figure 3: Abrogation of ERK1/2IEC results in enhanced ERK5 signalling.
(a,b) Ki67 staining of paraffin-embedded ileum sections from fl/fl and ΔIEC mice, 10 days after tamoxifen treatment. Nuclear staining with Hoechst. Sections were analysed by confocal fluorescent imaging; scale bar, 100 μm (a). Enumeration of the number of Ki67-positive cells per crypt (b). Data are mean±s.e.m. of 10 crypts (n=3 per group). **P<0.001 (Student’s t-test). (c) Immunoblotting for indicated proteins with IEC lysates isolated from fl/fl and ΔIEC mice as in a. (d) Immunoblotting for indicated proteins with IEC lysates as in a. (e) HCT116 cells were treated with the specific MEK1/2 inhibitor, PD0325901, for 0, 1, 2, 4 or 6 h and analysed by immunoblotting for indicated proteins. (f) HCT116 cells were treated with PD0325901 (0, 1, 10 and 100 nM) for 6 h and analysed by immunoblotting for indicated proteins. As control, some cells were co-treated with the specific ERK5 inhibitor, XMD8-92 (10 μM). (g) DLD-1 cells were treated as in e, and cell lysates analysed by immunoblotting. (h) DLD-1 cells were treated as in f, and lysates were analysed by immunoblotting. (i) Caco2 cells were treated as in e, and lysates were analysed by immunoblotting. (j) Caco2 cells were treated as in f, and analysed by immunoblotting. All data are representative of two to three experiments.
