Figure 5.

Interaction of ELMO1 and bacterial effectors with WxxxE signature motif controls inflammatory responses (a) (upper panel) Recombinant His-ELMO1-full length (FL) was incubated with GST, GST-IpgB1, GST-IpgB2, or GST-Map immobilized to glutathione sepharoseaffinity beads. The bound ELMO1-FL was visualized by immunoblot . Equal loading of GST proteins was confirmed by Ponceau S staining . (lower panel) The input of His ELMO-1- FL was shown. (b) Control (C1) or ELMO1 (E1) shRNA J774 cells were transfected with GFP vector control, GFP-SifA, GFP-IpgB1, GFP-IpgB2, and GFP-Map. The cell lysates were assessed by anti-GFP antibody (upper panel), and anti-ELMO1 antibody (middle panel). Equal loading was confirmed by α-Tubulin (lower panel). (c-e) Control or ELMO1 shRNA J774 cells were transfected with GFP vector, GFP-SifA, GFP-IpgB1, GFP -IpgB2, and GFP-Map as in (B), and then the cells were challenged with E.coli K12 (c) or LPS (d-e). The supernatants were collected and the level of TNF-α (c, d) and IL-6 (e) was measured by ELISA. Data in C-E represent mean ± SEM of three independent experiments. * means p ≤ 0.05 as assessed by one-way ANOVA multiple comparisons