Fig. 1: A Shigella-derived trans-acting factor antagonizes LPS ubiquitylation by RNF213.
From: Shigella flexneri evades LPS ubiquitylation through IpaH1.4-mediated degradation of RNF213
a, Representative confocal micrographs of HeLa cells at 5 h after infection with Salmonella Typhimurium (top) or S. flexneri (bottom), immunostained for LPS (red) and conjugated ubiquitin (FK2) (green) or LPS (red) and RNF213 (green) as indicated. Dashed squares, representative areas magnified ×2.5 in the insets. Scale bar, 10 μm. The micrographs are representative of n = 3 experiments. b, Immunoblot analysis of the indicated strains of Salmonella Typhimurium and S. flexneri extracted from HeLa cells at 5 h after infection. Gray triangle, ΔrfaL LPS; black circle, ubiquitin; GroEL, loading control for bacterial lysates. The LPS fractions were isolated by heat clearance of bacterial lysates and probed for conjugated ubiquitin with FK2 antibody. The loading control (GroEL) was probed for in non-heat-cleared bacterial lysates. The results are representative of n = 3 experiments. c, Percentage of bacteria positive for conjugated ubiquitin (detected by FK2 antibody) (left) and RNF213 (right) at 4 h after infection in HeLa cells. Data are averaged from n = 2 experiments for singly infected cells or n = 4 experiments for the double infection.
