Extended Data Figure 8: C3 modification/inactivation of the Rho subfamily induces Pyrin inflammasome activation.
From: Innate immune sensing of bacterial modifications of Rho GTPases by the Pyrin inflammasome
C3 toxin was delivered into indicated BMDMs in a, c–e or DC2.4 cells stably expressing Flag–RHOA/Cdc42 in b by using the LFn-PA system. The SDS–PAGE mobility shift in anti-RHOA/Cdc42 immunoblots in a shows the specific modification of RHOA by C3. Flag–RHOA/Cdc42 purified from DC2.4 cells was subjected to TcdB glucosylation in the presence of UDP-[3H]Glucose. 3H-autoradiograph and anti-Flag immunoblot in b show that C3 stimulation blocks TcdB modification of RHOA but not Cdc42. Caspase 1 immunoblots are shown in c; ELISA assay of IL-1β release in LPS-primed BMDMs is in d and e (n = 3; mean ± s.d.). Data in all panels are representative of at least three repetitions.
