Fig. 3.
Loss of ADAM17 abrogates EGFR signaling in colonic epithelia. a. Immunoblot of EGFR and pEGFR in the distal colon from control and Adam17flox/floxMx1-Cre+ (ΔAdam17_Mx1) mice at days 0 and 8 after DSS administration. β-Actin was used as a loading control. b. Phosphorylated EGFR (pEGFR), EGFR and TGF-α immunostaining of representative colon sections from control and ΔAdam17_Mx1 mice at day 8 after DSS administration. NI, non-immune IgG (Control for anti-pEGFR and anti-EGFR antibodies). Scale bars, 100 μm. c. Morphology of isolated colonic crypts from control and ΔAdam17_Mx1 mice at days 0 and 8 after DSS administration. Arrows indicate regenerative large crypts. Scale bars, 50 μm. d. Immunoblot of ADAM17, pEGFR and EGFR in isolated colonic crypts from control and ΔAdam17_Mx1 mice at day 8 after DSS administration. β-Actin was used as a loading control. e. Relative gene expression of MUC2 by RT-qPCR in isolated colonic crypts from control and ΔAdam17_Mx1 mice at day 8 after DSS administration. Bars, mean ± s.d.; n = 4 independently isolated colonic crypts; **, p < 0.01. f. Dot immunoblot of MUC2 in isolated colonic crypts from control and ΔAdam17_Mx1 mice at day 8 after DSS administration. Two μg of tissue lysates were loaded per dot and GAPDH was used as a loading control. Results between the two independent groups were determined by Student's t-test. P values smaller than 0.05 are indicated on respective plots.