Igf 1r sc 713

Immunoblotting was performed as described [6] (link), employing antibodies to phosphotyrosine (4G10, 05-321, EMD Millipore), PTEN (#9552), P-Smad 1/5/8 (#9511), P-Smad 3 (#9520), P-PKC[T514] (#9379), P-PKC[T638/641] (#9375), P-PKC[S660] (#9371), P-S6 (#2211), P-Akt[S473] (#9271), P-Akt[T308] (#9275), P-GSK3[S9] (#9331), and GSK3 (#9315) from Cell Signaling Technology (Danvers, MA) or IGF-1R (sc-713), and E2F4 (sc-866) from Santa Cruz Biotechnology (Santa Cruz, CA). All other antibodies used were purchased from commercial sources and listed previously [11] (link)–[13] (link).

The following primary antibodies were used in this study for immunoblotting: pRB^ser780 (CST-3590), pRB^ser807 (CST-8516), total-RB (CST-9309), cyclin D1 (CST-2922), cyclin D3 (CST-2936), pAKT^ser473 (CST-9271), pAKT^Thr308 (CST-9275), total-AKT (CST-9272), pEGFR^Tyr1068 (CST-3777), total-EGFR (CST-2232), pERBB2^Tyr1248 (CST-2243), total-ERBB2 (CST-4290), pERBB3^Tyr1222 (CST-4784), pIGF1R^Tyr1135 (CST-3918), pS6K^Ser235/236 (CST-2211), total-S6K (CST-2217), Raptor (CST-2280), RheB (CST-13879), p4EBP1^Thr37/46 (CST-2855), 4EBP1 (CST-9452), pSIN1^Thr86 (CST-14716), SIN1 (CST-12860), pER^ser167 (CST-5587), Rictor (CST-2114) and Deptor (SCT-11816) were purchased from Cell Signaling Technology. p107 (sc-318), p130 (sc-317), total-ER (sc-8002, F-10), ERBB3 (sc-415) and IGF1R (sc-713) were purchased from Santa Cruz Biotechnology. β-tubulin (T-9026) were from Sigma-Aldrich and Ki67 from Clinisciences. The following antibodies were used for immunohistochemistry: pERK1/2^Thr202/4 (CST-4370), pAKT^ser473 (CST-4060), pS6K^Ser235/6 (CST-4858), pmTOR^Ser2448 (CST-2976) and p4EBP1^Thr37/46 (CST-2855) were purchased from Cell Signaling Technology. Ki67 was purchased from Clinisciences. Reagents were obtained from the following sources: 17-β-oestradiol (E2) and 4-hydroxytamoxifen (4-OHT) from Sigma-Aldrich, fulvestrant from Tocris, and neratinib and vistusertib from SelleckChem.