Rabbit anti phospho eif2α 119a11

Western blotting was performed as described previously (Florentin et al., 2017). Parasite pellets were isolated using cold 0.04% Saponin (Sigma) in 1X PBS for 10 min as described previously (Florentin et al., 2017; Muralidharan et al., 2011). Antibodies used for this study were the following: mouse anti‐GFP JL‐8 (Clontech, 1:3000), rabbit anti‐PfEF1α (from D. Goldberg, 1:2,000), mouse anti‐plasmepsin V (from D. Goldberg, 1:400), rabbit anti‐PfBiP MRA‐1246 (BEI resources, 1:500), rabbit anti‐GFP A‐6455 (Invitrogen, 1:2,000), mouse anti‐eIF2α L57A5 (Cell Signalling, 1:1,000), rabbit anti‐Phospho‐eIF2α 119A11 (Cell Signalling, 1:1,000), rat anti‐HA (Roche 3F10, 1:3000), mouse anti‐Ty1 (Sigma Clone BB2, 1:1000), and mouse anti‐Ub P4D1 (Santa Cruz Biotechnology, 1:1,000). Secondary antibodies used were IRDye 680CW goat anti‐rabbit IgG and IRDye 800CW goat anti‐mouse IgG (LICOR Biosciences, 1:20,000). The western blots were imaged using the Odyssey infrared imaging system. Polyacrylamide gels used in this study were either prepared using 10% EZ‐Run protein gel solution (Fisher) or precast gradient gels (4–20%, from Bio‐Rad). Any quantification performed on western blots was done using ImageJ software. The quantification data were analysed using Prism (GraphPad Software, Inc.).