Protein extract preparation and immunoblotting were performed accordingly to standard procedures [36 (link),38 (link)]. The Abs used for the study were as follows: anti-PI3Kα (#4249S), anti-PI3Kβ (#3011S), anti-PI3Kδ (#34050S), anti-p-STAT3 (Tyr705#9131S), anti-p-AKT (T308#9275S and S473#9271S), anti-p-PDK1 (S241#3061S), anti-PDK1 (#3062S), anti-p-S6 (S235/236#2211), anti-S6 (#2317S), and anti-p-p65 (S276#3033S) (all from cell signaling); anti-K10 (#PRB-159P) and anti-Loricrin (PRB-145P) (both from Covance); anti-cyclin D1 (#sc-20044), anti-STAT3 (C-20#sc-482), and anti-β-actin (all from Santa Cruz Biotechnology, Santa Cruz, CA, USA); and anti-keratin 5 (K5) (#MA5-14473, Invitrogen). Filters were properly developed with anti-mouse, anti-goat, or anti-rabbit Ig Abs conjugated to HRP using the ECL-plus detection system (Amersham, Dubendorf, Switzerland), or, otherwise, the SuperSignal West Femto kit (Pierce, Rockford, IL, USA). Immunoblots were subjected to densitometry using the ChemiDoc MP Imaging System (Bio-Rad, Hercules, CA, USA) supported by the Molecular Analyst software (https://imagej.nih.gov/ij/, accessed on 20 July 2021). Band intensities were evaluated in three independent experiments and reported as means of densitometric intensity (D.I.) ± SD.
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