Comprehensive Western Blot Analysis of Key Signaling Pathways

Western blot was conducted as per our previous reports^{6 (link),25 (link),48 (link)}. Briefly, blots were first incubated with antibodies to the following proteins: Nrg4 (1:1,000 dilution, Thermo Fisher, PA5-102641), P-IKKβ (Ser176/180) (1:1,000 dilution, CST, 2697), IKKβ (1:1,000 dilution, CST, 2684), P-p65 (Ser468) (1:1,000 dilution, CST, 3039), p65 (1:1,000 dilution, CST, 8242), P-IκBα (Ser32) (1:1,000 dilution, CST, 2859), IκBα (1:1,000 dilution, CST, 9242), P-ERK (Thr202/Tyr204) (1:1,000 dilution, CST, 4376), ERK (1:1,000 dilution, CST, 4695), P-Akt (Ser473) (1:1,000 dilution, CST, 4051), Akt (1:1,000 dilution, CST, 4691), AMPK (1:1,000 dilution, Abcam, ab32047), P-AMPK (Thr183 + Thr172) (1:1,000 dilution, Abcam, ab133448), ErbB4 (1:1,000 dilution, Abcam, E200), MMP2 (1:1,000 dilution, Abcam, ab92536), MMP9 (1:1,000 dilution, Abcam, ab76003) and GAPDH (1:3,000 dilution, CST, 5174). Binding of the primary antibody was detected by incubating membranes with a horseradish peroxidase-conjugated goat anti-mouse secondary antibody (1:2,000 dilution, Abcam, ab205719).