Anti ef2 c 14

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.

Anti‐Rab5C (Ab199530; Abcam, Cambridge, UK), Anti‐red, for detection of mCherry, (5F8 Chromotek, Munich, Germany), Anti‐ATP6V1G1 (16143‐1‐AP; Proteintech Europe, Manchester, UK), Anti‐EF2 (C14; Santa Cruz, Dallas), Anti Lamp‐1 (H4A3; Abcam), Alexafluor 568 anti‐rat (Thermo Fisher, Waltham). Horseradish peroxidase (HRP)‐conjugated secondary antibodies (Sigma Aldrich).