Tubulin α clone dm1a

Cell lysate was subjected to sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. The protein was transferred to a nitrocellulose membrane and hybridized with antibody against AMPKα (catalog number: #2532), phosphorylated AMPKα at Thr 172 (#2535), 4E‐BP1 (#9452), phosphorylated 4E‐BP1 at Thr 37/46 (#9459), p70S6K (#9202), phosphorylated p70S6K at Thr389 (#9205), phosphorylated p53 at Ser 15 (#9284), AKT (#9272), phosphorylated AKT at Ser 473 (#9271), ACC at Ser 79 (#3661), actin (#4970) (Cell Signaling, Danvers, MA, USA), phosphorylated H2AX at Ser 139 (#613401) (BioLegend, San Diego, CA, USA), p53 (Ab‐6, clone DO‐1) and tubulin‐α (clone DM1A, Thermo Fisher Scientific, Fremont, CA, USA) followed by an appropriate second antibody. The membranes were developed with the ECL system (GE Healthcare, Buckinghamshire, UK). actin and tubulin‐α were used as a loading control. DMSO, a solvent of nutlin‐3a, was used as a control.

Cell lysate was subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The protein was transferred to a nitrocellulose membrane and was hybridized with antibody against integrin β-3 (Catalog Number: #13166), AMPK (#2532), phosphorylated AMPK (Thr 172) (#2535), phosphorylated p53 (Ser 15) (#9284) (Cell Signaling, Danvers, MA, USA), plasminogen activator inhibitor (#ab20562), a disintegrin and metalloproteinase with thrombospondin motifs 5 (#ab41037) (Abcam), IGFBP3 (#sc-9028), CARP (sc-30181), β-2 adrenergic receptor (#sc-569) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), phosphorylated H2AX (Ser 139) (#613401) (BioLegend, San Diego, CA, USA), p53 (Ab-6, Clone DO-1) and tubulin-α (Clone DM1A) (Thermo Fisher Scientific) as a control followed by an appropriate second antibody. The membranes were developed with the ECL system (GE Healthcare, Buckinghamshire, UK).