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. Author manuscript; available in PMC: 2019 Apr 2.
Published in final edited form as: Sci Signal. 2018 Oct 2;11(550):eaat8335. doi: 10.1126/scisignal.aat8335

Fig. 8. Glu3930 and His4030 are essential for RRSP cytotoxicity.

Fig. 8.

(A, B) Representative immunoblots of lysates from HeLa cells incubated with LFNRRSP, the LFNRRSP E3930A and H4030A mutants, and anthrax toxin protective antigen (PA) as indicated and probed with antibodies that recognize all RAS proteins (Pan-RAS) (A) or total ERK1/2 and phosphorylated ERK1/2 (pERK1/2) (B). (N=3 independent experiments) (C) Percentage of RAS that was cleaved in HeLa cells by wild-type and mutant LFNRRSP as determined by densitometry analysis. (D) The ratio of pERK1/2 to total ERK1/2 in lysates from HeLa cells following treatment with wild-type and mutant LFNRRSP. (E) Schematic representation of MARTX toxin effector domain organization in V. vulnificus CMCP6 strains. (F) Representative immunoblots showing RAS in lysates from T84 cells that has been incubated with V. vulnificus producing the indicated MARTX toxins (N=3). (G) Percentage of Ras that was cleaved in (E) as determined by densitometry analysis. All graphs (C, D, G) show means ± SD from three independent biological replicates (ANOVA followed by Dunnett’s multiple comparisons test; *P≤0.05, **P≤0.01 and ****P≤0.0001). n.d., not detected.