Her3 erbb3

Anti-MSI2 (#ab76148), anti-MSI1 (#ab21628), and anti-β-actin HRP conjugated (#ab49900) antibodies and recombinant MSI2 protein (#ab167853) were obtained from Abcam, (Cambridge, UK). Anti-EGF receptor (#4267), phospho-EGFR (Y1068) (mAb #2234), HER3/ErbB3 (#12708), HER2/ErbB2 (#4290), Smad3 (#9523), phospho-AKT (T308) (#13038), total AKT (#2920), phospho-ERK (T202/Y204) (#4370), total ERK (#4696), phospho-p70S6K (T389) (#9234), total p70S6K (#2708), and normal Rabbit IgG (#2729) were obtained from Cell Signaling, (Danvers, MA). Erlotinib and afatinib were obtained from LC Laboratories (Woburn, MA), doxycycline from Sigma-Aldrich (D9891, Darmstadt, Germany). SUPERase-In RNAse inhibitor was obtained from Thermo Fisher Scientific, (AM2694 Waltham, MA). Osimertinib was obtained from Selleckchem (#S7297, Houston, TX)

Cells were lysed in RIPA buffer. Equal amounts of total protein were resolved by SDS-PAGE, transferred to PVDF membranes (Millipore), and immunoblotted with the indicated primary antibodies. Membranes were hybridized with the following primary antibodies: p-Erk1/2, Erk1/2, p-HER3/ErbB3, HER3/ErbB3, HER2/ErbB2, p-AKT, AKT (Cell Signaling Technology), NIS, Tg, TPO, TSHR, GLUT1, and GLUT3 (Protein tech). Membranes were then hybridized with species-specific HRP-conjugated antibodies. Bands were visualized with Potent ECL kit (Beyotime).