Ptbc1d1 ser237

Unless otherwise stated all reagents were from Sigma–Aldrich. Primary antibodies against pan-AMPK-α (#2532), AMPK-β1/2 (#4150), pAMPK Thr¹⁷² (#2531), AKT (#4685), ACC (#3876), pACC Ser⁷⁹ (#3661), IRAP (#3808), rabbit FLAG (#2368), Raptor (#2280), pRaptor Ser⁷⁹² (#2083) and TBC1D1 (#5929) were from Cell Signaling Technology. Other antibodies were pan-14-3-3 (sc-629) and GST (sc-138) from Santa Cruz Biotechnology Inc., GFP (118144600001; Roche), AMPK-γ1 (ab32508; Abcam), pTBC1D1 Ser²³⁷ (#07-2268; Merck-Millipore), β-actin (A1978) and mouse FLAG (F1804). Isoform-specific AMPK-α antibodies were described previously [34 (link)]. A769662 (Abcam) and AICAR (Tocris) were used.

Antibodies against AMPKα1/2, AMPKα2, p‐AMPKα1/2(Thr¹⁸³/Thr¹⁷²), phospho‐ACC (Ser⁷⁹), ACC, and 14‐3‐3 were obtained from Abcam (San Francisco, CA, USA). TBC1D1 and p‐TBC1D1 (Ser²³⁷) were purchased from Merck Millipore (Darmstadt, Germany). Antibodies against HDAC5 and p38 MAPK were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against β‐actin were purchased from Sigma‐Aldrich (St. Louis, MO, USA). Antibodies against phospho‐p38 (Thr¹⁸⁰/Tyr¹⁸²), α‐tubulin, GLUT4, and lamin B were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Phospho‐HDAC5 (Thr⁴⁹⁸) antibodies were from Thermo Fisher Scientific (Rockford, IL, USA). Horseradish peroxidase (HRP)‐conjugated goat anti‐rabbit IgG and goat anti‐mouse secondary antibodies were purchased from Enzo Life Sciences (Farmingdale, NY, USA). Metrnl was obtained from Cusabio (Wuhan, Hubei, China), and 1, 2‐bis (o‐aminophenoxy) ethane‐N, N, N′, N′‐tetraacetic acid (BAPTA)‐AM was purchased from Abcam. Compound C and STO‐609 were obtained from Calbiochem (San Diego, CA, USA). Protein A‐agarose beads were obtained from GE Healthcare (Piscataway, NJ, USA). The fluorescent Ca²⁺ indicator Fluo‐3 AM and Hoechst 33342 were obtained from Invitrogen (Leiden, the Netherlands).

Unless otherwise stated all reagents were from Sigma-Aldrich. Primary antibodies against pan-AMPK-α (#2532), pAMPK Thr¹⁷² (#2531), ACC (#3876), pACC Ser⁷⁹ (#3661), EGFR (#2232), IRAP (#3808), N-cadherin (#14215), Rab10 (#8127S) and rabbit FLAG (#2368) were from Cell Signaling Technology. CD36 (ab124515), Calmodulin (ab45689), GLUT1 (ab115730) and SERCA1 (ab129104) were from Abcam. From Santa Cruz Biotechnology Inc were antibodies against pan-14-3-3 (sc-629) and transferrin receptor (sc-65882). Other antibodies were EHBP1L1 (#PAB15793; Abnova), GFP (118144600001; Roche), HA.11 16B12 (901503; Biolegend), MICAL1 (14818–1-AP; proteintech), pTBC1D1 Ser²³⁷ (#07-2268; Merck-Millipore), VPS13C (HPA043356; Atlas Antibody), β actin (A1978), Tubulin (T9026) and mouse FLAG (F1804). VPS13A antibody was produced as previously described^{84 (link)}. Secondary antibodies used were conjugated with an AlexaFluor (Invitrogen) or HRP (Cell Signaling Technology or Jackson ImmunoResearch). AICAR was from Tocris.