Extended Data Fig. 4. SopD enhances pro-inflammatory signaling by antagonizing Rab8 through its GAP activity.

(a) Expression levels of SopD and its catalytic mutant SopD^R312A in stable cell lines (Raw264.7). Stable cell lines (Raw264.7) expressing HA-tagged SopD or SopD^R312A were lysed before immunoblotting analysis with the antibodies directed to the HA tag and to β-actin (as a loading control). (b) Effect of the expression of SopD or its catalytic mutant SopD^R312A on LPS-induced activation of AKT, p70S6K, Erk1/2, and p38 MAP, and NF-κB signaling pathways. Raw264.7 cells stably expressing HA-tagged SopD or its GAP-deficient mutant SopD^R312A were treated with LPS (100 ng/ml) for the indicated times, lysed, and analyzed by immunoblotting with antibodies specific for the phosphorylated state of AKT, p70S6K, p38, and Erk1/2, as well as an antibody to I-κBα and β-actin (as a loading control). The quantification of the western blot analyses is shown on Fig. 2e.