The Salmonella Type III Effector SspH2 Specifically Exploits the NLR Co-chaperone Activity of SGT1 to Subvert Immunity
Figure 5
S. Typhimurium SspH2 does not alter the essential cell cycle function of SGT1.
A, Dilution series of wild type yeast expressing empty vector (EV) or SspH2 with HA-epitope tag grown at 25°C. The expression of HA-tagged SspH2 was assessed by immunoblot in total lysate. B, Dilution series of yeast sgt1-3 complemented with ScSgt1p (Sc), HsSGT1A (HsA), HsSGT1B (HsB) or empty vector (EV) and expressing SspH2-HA (+) or empty vector (−), were grown at the permissive (25°C) and restrictive (32°C) temperatures. C, The expression of HA-tagged SspH2 in cultures from (B) was assessed by immunoblot in total cell lysate. The membrane was stained with Ponceau S (Pon. S) to indicate protein loading. D, HeLa cells transfected with plasmid control (EV) or constitutive expression vectors for SspH1 or SspH2 were quantified for cytotoxicity. Data are presented as the mean ± standard error of the mean for three independent determinations. S. Typhimurium effectors were tagged with an HA epitope and shown in the inset is the expression of transfected constructs as assessed by α-HA IB. Calnexin is a loading control.