Fig. 3: OspG resculpts host ubiquitination landscape.

a Immunofluorescent staining of HeLa cells transfected with GFP and GFP-OspG for 24 h. The ubiquitinated proteins were stained with the FK2 antibody, and nuclei were stained with DAPI. Fluorescence intensity was plotted along the arrows. Scale bar, 20 μm. Images are representative of n  =  2 independent experiments. b Schematic diagram of the ubiquitome profiling under S. flexneri infection. HeLa cells were infected with indicated S. flexneri strains, and the NHS-activated agaroses coupled with TUBE were used to capture ubiquitinated proteins. TUBE, tandem-repeated ubiquitin-binding entities. RP-LC, reversed-phase liquid chromatography. c Immunoblot for ubiquitin (P4D1 antibody) showing TUBE capture of ubiquitinated proteins from cell lysates as in (b). Images are representative of n  =  3 independent experiments. Input, cell lysates before TUBE capture. Output 1 and output 2, supernatant of cell lysates after the first and the second TUBE enrichment, respectively. Ub, ubiquitin.