Fig. 4: The Salmonella effector SopF blocks ER-phagy.
From: The ER-phagy receptor FAM134B is targeted by Salmonella Typhimurium to promote infection
A WT HEK293A cells expressing ss-RFP-GFP-KDEL were AA-starved for 6 h or 3 h, then infected with ST WT or ST ΔsopF for 3 h. ER-phagy was measured via ss-RFP-GFP-KDEL processing. Actin = loading control. NR nutrient rich; AA amino acids; ST = Salmonella Typhimurium. B Normalized Free RFP-Actin ratio from (A). Data are presented as mean values ± SD of five biological experiments. ANOVA, ***P < 0.001; ns no significance. C WT HEK293A cells expressing ss-RFP-GFP-KDEL were AA-starved for 6 h or 3 h + ST WT/ΔsopF infection for 3 h, fixed, and imaged. ss-RFP-GFP-KDEL = yellow; Free RFP = red. Scale bar = 5 μm. D Average RFP puncta/cell from (C). Data are presented as mean values ± SD of five biological experiments. ANOVA, ***P < 0.001. E WT cells from (A) transfected with FLAG-SopF or mock plasmid, AA-starved for 6 h or 3 h + ST WT infection for 3 h. ER-phagy measured. Actin = loading control. F Normalized Free RFP-Actin ratio from (E). Data are presented as mean values ± SD of three biological experiments. ANOVA, ***P < 0.001. G HEK293A FAM134B KO cells with ss-RFP-GFP-KDEL transfected with FAM134B WT, G216R, or FLAG-SopF, then AA-starved for 6 h. Vinculin = loading control. H Normalized Free RFP-Vinculin ratio from (G). Data are presented as mean values ± SD of five biological experiments. ANOVA, **P < 0.01; ***P < 0.001.
