Anti α tubulin t6199 200ul

For Western blotting, anti-Flag (M2) (F3165-5MG, 1:5000 dilution) and anti-α-tubulin (T6199–200UL, 1:5000 dilution) monoclonal antibodies were obtained from Sigma-Aldrich. An anti-Myc (sc-40, 1:500 dilution) monoclonal antibody was purchased from Santa Cruz Biotechnology. An anti-hemagglutinin (HA) monoclonal antibody (MMS-101P, 1:3000 dilution) was obtained from BioLegend. Anti-PJA2 (#40180, 1:1000 dilution), anti-MOB1 (# 3863S, 1:2000 dilution), and anti-phospho-p70 S6 kinase (Thr389) (#9234S,1:1000 dilution) polyclonal antibodies were obtained from Cell Signaling Technology. An anti-S1P (Z-P300; 1:500 dilution) monoclonal antibody was obtained from Echelon Biosciences.
For immunostaining, anti-HA (3724S, 1:3000 dilution) and anti-LAMP1 (9091S, 1:400 dilution) polyclonal antibodies were obtained from Cell Signaling Technology. An anti-Flag (F7425-.2MG, 1:5000 dilution) polyclonal antibody was purchased from Sigma-Aldrich. An anti-TOM20 (612278, 1:200 dilution) monoclonal antibody was purchased from BD Biosciences.
For chemicals, FIPI hydrochloride hydrate (#F5807) was obtained from Sigma-Aldrich. CAY10594 (#13207) was obtained from Cayman Chemical.

For pull-down assays, 1 × 10⁷ cells were lysed with NETN buffer containing protease inhibitors on ice for 20 min. Cell lysates were collected after centrifugation and incubated with 20 μL of S-beads for 2 h at 4 °C. The beads were washed with NETN buffer three times and boiled in 2× Laemmli buffer. The samples were resolved using SDS–polyacrylamide gel electrophoresis and transferred to a polyvinylidene fluoride membrane; immunoblotting was carried out with antibodies, as indicated in the figures.
Anti-ERK1/2 (9102, CST), anti-pERK1/2 (9101, CST), p-Ser/Thr (9631, CST) and anti-eIF5A (20765, CST) antibodies were purchased from Cell Signaling Technology and used at 1:1000 dilution. Anti-DHPS (NBP2-32256, Nova Biomedical) was used at 1:2000; anti-α-tubulin (T6199-200UL) and anti-Flag (M2) (F3165-5MG) monoclonal antibodies were purchased from Sigma-Aldrich and used at 1:5000 dilution. Anti-Hypusine antibody (ABS1064, Sigma) was used at 1:500 dilution.