Protein Expression Analysis by Western Blot

Protein lysates were collected and analyzed by western blot as previously described [42 (link)]. In brief, primary antibodies against p90RSK (Ser380) (Cell Signaling Technology, Danvers, MA, USA, 9341), p70 S6 Kinase (Thr389) (Cell Signaling Technology 9205), S6 ribosomal protein (Ser235/236) (Cell Signaling Technology, 4856), eIF4G (Ser1108) (Cell Signaling Technology, 2441), RVFV MP12 Antibody (IBT Bioservices, Rockville, MD, USA, 04-0001), or HRP-conjugated actin (Abcam, ab49900) were diluted 1:1000 in 5% bovine serum albumin in 1× TBS with 0.1% Tween-20 solution followed by the addition of the appropriate secondary antibody. The western blots were visualized by chemiluminescence using SuperSignal West Femto Maximum Sensitivity Substrate kit (ThermoScientific, Waltham, MA, USA) and a Bio Rad Molecular Imager ChemiDoc XRS system (Bio-Rad, Hercules, CA, USA) [41 (link),42 (link)].