Fig 4.

Impact of PPP inhibition with CA and Yptb infection on pyrin oligomers and phosphorylation as determined by BN-PAGE and immunoblotting. WT BMDMs left untreated (A and B) or pretreated and maintained with DMSO or 10 nM CA (C–E) were left uninfected or infected with WT or ∆yopM Yptb at an MOI of 30 for 90 min. BMDM lysate samples prepared in 1% digitonin were separated by BN-PAGE and immunoblotted. Insoluble proteins from 1% digitonin lysates of DMSO-treated samples were analyzed by SDS-PAGE and total pyrin immunoblotting (C). BN-PAGE MW standards are shown on the left, and tentative positions of pyrin oligomers are shown on the right. Dashed horizontal lines in (D and E) aid viewer appreciation of band upshifts in the presence of CA.