Western blot analysis of extracts of HeLa cells, using Phospho-EIF4B-S422 antibody at 1:2000 dilution. HeLa cells were treated by EGF (100ng/ml) for 30 minutes after serum-starvation overnight. The secondary antibody used was an HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates were loaded 25ug per lane and 3% nonfat dry milk in TBST was used for blocking. Blocking buffer: 3% BSA.An ECL Kit was used for detection and the exposure time was 60s.