Fig. 3: FAM134B oligomerization is hindered by Salmonella infection.

A FAM134B KO HEK293A cells transfected with FAM134B-FLAG were starved for AA for 3 h in the absence or presence of ST. FLAG was immunoprecipitated. Data are presented as mean values ± SD of five biological experiments. Student’s t test, ***P < 0.001. AA amino acids; ST, Salmonella Typhimurium. B WT cells were incubated with BafA1 and starved for AA for 3 h in the presence or absence of ST. Fam134B puncta formation was observed by confocal microscopy. NR, Nutrient Rich; BafA1, Bafilomycin A1. Data are presented as mean values ± SD of five biological experiments. The average number of Fam134B puncta was calculated from a minimum of 100 cells. Representative images are shown. Fam134B, red; DAPI, blue; ST, green. Scale bar 5 μm. ANOVA, *** P < 0.001. C FAM134B KO cells were transfected with either FAM134B WT or FAM134B G216R and starved for AA for 6 h or starved for 3 h, followed by ST infection for 3 h in AA starvation media. ER-phagy was measured by ss-RFP-GFP-KDEL processing. Actin was used as a loading control. D Normalized Free RFP-Actin ratio from cells in (C). Data are presented as mean values ± SD of five biological experiments. ANOVA, ***P < 0.001; ns, no significance. E FAM134B KO were transfected with either FAM134B WT or G216R and starved for AA for 6 h or starved for 3 h followed by ST infection for 3 h in AA starvation media. Representative images are shown. ss-RFP-GFP-KDEL, yellow; Free RFP, red; Fam134B, far red. Scale bar 5 μm. F Average number of RFP puncta per cell from (E) were quantified. Data are presented as mean values ± SD of five biological experiments. The average number of RFP punctuation was calculated from a minimum of 100 cells. ANOVA, ***P < 0.001. Source data are provided as a Source Data file.