Anti eif4ebp1

Western blot analysis was performed as described.[21 (link)] The primary antibodies used in this study were anti-pMTOR (Ser2448), anti-E2F2, anti-SLC2A2, anti-SLC2A3, and anti-SLC2A4 (Abcam, Cambridge, UK), anti-YY1, and anti-CCNA2 (Santa Cruz Biotechnology), anti-MYC, anti-CCNE1, anti-EIF4EBP1, anti-pEIF4EBP1 (Thr37/46), anti-RPS6KB1, and anti-pRPS6KB1 (Thr389) (Cell Signaling Technology, Danvers, MA), anti-SLC2A1 (Novus Biologicals, Littleton, CO), anti-HA-tag (Zymed Laboratories, South San Francisco, CA), and anti-ACTB (actin, beta) (Chemicon, Temecula, CA). The plasma membrane proteins were prepared with the Qproteome Plasma Membrane Protein Kit (Qiagen, Valencia, CA) following the manufacturer’s instructions.

Antibodies used in this study include: anti-U2AF1 (ab86305, Abcam), anti-U2AF2 (sc-48804, Santa Cruz Biotechnology), anti-hnRNP A1 (4B10, Abcam), anti-RPS6 (#2317, Cell Signaling), and anti-pRPS6 (#2211, Cell Signaling), anti-TUBULIN (sc-53646, Santa Cruz Biotechnology), anti-SRSF3 (sc-13510, Santa Cruz Biotechnology), anti-EIF4EBP1 (#9452, Cell Signaling), anti-HNRNPC1/C2 (ab10294, Abcam), anti-Flag (F3165, Sigma Aldrich).

Phosphorylated and total forms of proteins were detected by immunocytochemistry using antibodies from the following companies: anti-phospho-eIF4E-BP1 (Ser65) (1:100), anti-eIF4E-BP1 (1:100), anti-mTOR (1:100), anti-phospho-Akt (Ser473) (1:250), anti-Akt (1:250), anti-phospho-eEF2 (Thr 56) (1:100), anti-eEF2 kinase (1:100), and anti-phospho-eEF2 kinase (Ser366) from Cell Signaling Technology, Danvers, MA, USA; anti-phospho-mTOR (S2448) (1:250) from Abcam, Cambridge, MA, USA; and anti-EF2 (C-14) (1:100) from Santa Cruz Biotechnology, Santa Cruz, CA, USA. F-actin was visualized with Alexa-Fluor 594-phalloidin (used at 1:50; Invitrogen Life Technologies, Carlsbad, CA, USA), and β-tubulin was labeled with anti-β-tubulin (used at 1:1000; Sigma-Aldrich Corporation, St. Louis, MO, USA). Secondary antibodies conjugated with Alexa Fluor dyes were from Invitrogen Life Technologies, Carlsbad, CA, USA.