(A-C) Levels of phosphorylated (p)-EGFR and p-ERK, HB-EGF, EGF, precursor (pre)-EGF, and pre-HB-EGF in the epidermis of WT (W) and IkkαF/F/K5.Cre (M) mice, as detected by Western blotting. P, postnatal day; β-Tubulin, loading control.
(D) mRNA levels of ADAMs in WT and Ikkα−/− (−/−) primary keratinocytes cultured with no growth supplement for 1 day (Starv) or with growth supplement (Supplem), as detected by RT-PCR. GAPDH, PCR control.
(E) Levels of ADAM12, 10, and 17 in the epidermis of WT and IkkαF/F/K5.Cre (M) mice, as detected by Western blotting.
(F) Activities of sheddases in the epidermis of WT, IkkαF/F/K5.Cre (M) and IkkαF/F/K5.Cre/K5.IKKα mice (M+IKKα), as detected by gelatin zymography. WB, Western blotting; *, indicates proteins.
(G) Levels of p-EGFR, p-ERK, EGF, HB-EGF, and/or ADAMs in WT, Ikkα−/− (−/−), and Ikkα−/−keratinocytes treated with GM6001 (GM), GW2974 (GW), and/or PD98059 (PD) for 24h or with reintroduced IKKα or IKKα-KA after 48 h, as detected by Western blotting. β-Actin, loading control.
(H) Levels of indicated proteins in the skin of WT, Ikkα−/− (−/−), Ikkα−/−/K5.IKKα newborns, as detected by Western blotting. NS, non-specific band.