Laminin a c

Recombinant human cytokines and anti-human IL-36γ were purchased from R&D Systems (Minneapolis, MN). Inhibitors of p42/44 MAPK (PD98059 and U0216) and inhibitor for p38 MAPK (SB203580) were purchased from Merck (Darmstadt, Germany). siRNA for NF-κB p65, c-Jun and a control siRNA were purchased from Santa Cruz (Santa Cruz, CA). Antibodies against phosphorylated and total p42/44 MAPK (ERK1/2), p38 MAPK, JNK1/2, GAPDH and laminin A/C were purchased from Cell Signaling Technology (Beverly, MA). Antibodies against phosphorylated c-Jun, NF-κB p65, phosphorylated IκBα were purchased from Santa Cruz. All other reagents were purchased from Sigma Chemical Co. (St Louis, MO).

2-deoxyglucose (2-DG, Sigma, D6134-250MG) was dissolved in 1x DPBS. SBI-0206965 (Cayman Chemical, 18477) and MK-8722 (Aobious, AOB33226) were dissolved in DMSO (Sigma-Aldrich). Hoescht 33342 (Cell Signaling, 4082S) and puromycin (Sigma-Aldrich, P9620) were dissolved in deionized water. For western blotting and immunostaining, antibodies from Cell Signaling Technologies (Denvers, MA USA) were used at 1:1000 dilution unless otherwise noted: P-ACC S79 (3661), P-AMPK T172 (2535), ACC (3662), AMPKα (2532), GAPDH (5174, 1:10,000), Laminin A/C (4777), LKB1 (3047), β-tubulin (2146, 1:10,000). From Sigma-Aldrich, anti-Actin (A5441) was diluted 1:10,000 and anti-Tubulin (T5168) was diluted 1:5000. From Santa Cruz, CaMKK2 (sc-100364) was diluted 1:500–1:1000. From Pierce, horseradish peroxidase-labeled goat anti-rabbit (PI31460) or anti-mouse (PI31430) was diluted 1:1000–1:10,000. Lipofectamine 2000 (11668019) was purchased from Thermo Fisher, and FuGENE HD (E2311) was purchased from Promega. MitoTracker Red (M22425) and LysoTracker Red (L7528) were purchased from Thermo Fisher and diluted in DMSO.