Skip to main content
. 2013 Mar 27;8(3):e60499. doi: 10.1371/journal.pone.0060499

Figure 2. Pore forming activities of SipC N-terminal mutant strains.

Figure 2

(A) Schematic diagram of the SipC N-terminus showing the deletion/insertion sites in SipC. (B) Sheep Red Blood cells were infected with various Salmonella strains and the relative hemolysis activity was measured at OD 595 nm. The hemolysis experiments were repeated three times with the standard errors shown. Statistical analysis of hemolysis was performed by Student's t test with a p value of <0.05 being considered significant (*). When compared to the wild-type strain, the p values of sipB, sipC, sipD, invA, sipC-N1, N2, N3, N4, N5, N6 mutant strains and the un-infected are 0.0005(***), 0.0026(**), 0.0002(***), 0.0003(***), 0.0009(***), 0.4872, 0.7540, 0.4709, 0.0124(*), 0.2500, 0.0001(***) respectively. The expression (C) and secretion (D) of SipC, SipB, SipD, SipA, and DnaK in Salmonella strains including the wild-type, sipB, sipC, sipD, invA, sipC-N1-N6. Bacterial strains were grown under SPI-1 inducing conditions and equal amounts of bacterial lysates or culture supernatants were analyzed by Western blotting using rabbit polyclonal anti-SipB, anti-SipC, anti-SipD, anti-SipA, monoclonal anti-M45 and anti-DnaK antibodies. (D) SipC-N5 is defective in binding to SicA. The interactions of SipC-N, SipC-N1-N6 with SicA were analyzed by pull-down assay using purified GST-SicA or GST as negative control. The presence of GST-SicA and His-SipC after pull down was determined by Western blotting with polyclonal anti-GST and monoclonal anti-His antibodies.