Fig. 1: High-throughput ADP-ribosylome profiling identifies septins as host targets of OspC effectors.
From: Shigella flexneri evades septin-mediated cell-autonomous immunity via protein ADP-riboxanation
a Identification of candidate substrates. Top: Venn diagram showing the overlap of OspC1 and OspC3 candidates. Bottom: Lysates from 293T cells transfected with WT OspC or its catalytically dead mutant (EH/AA) were probed with an anti-ADP-ribose antibody. b GO enrichment analysis. Dot plot of significantly over-represented GO terms. Adjusted P values (one-sided hypergeometric test, Benjamini-Hochberg correction) are color-coded. Dot sizes indicate fold enrichment. Non-significant terms are excluded. c Heatmap of conserved substrates. Conserved candidates of OspC1 and OspC3 are shown, highlighting SEPT2, SEPT7, and SEPT9. Data represent two biological replicates. d Immunoblotting analyses of a panel of septin subunits (SEPT2, 6, 7, 9, 10, 11) for their potential ADP-riboxanation. Immunoprecipitated septins from 293T cells co-expressing GFP-OspC3 or its EH/AA mutant were blotted with indicated antibodies. WCL, whole cell lysates. e–h SEPT2, SEPT6, SEPT7 and SEPT9 were modified by OspC1 and OspC2 as well. Immunoprecipitated septin samples from 293T cells co-transfected with OspC effectors were probed with the indicated antibodies. For (a, d–h), experiments were repeated three times with similar results.
