Volume 8 issue 10 October 2007
Bacteria-generated PtdIns(3)P recruits VAMP8 to facilitate phagocytosis
S Dai, Y Zhang, T Weimbs, MB Yaffe and D Zhou

Supplemental Figure 1: VAMP8 accumulation decrease when ruffles decrease following invasion. HeLa cells were infected for 10 minutes and bacteria were washed away followed by incubating in fresh media for 50 minutes in the absence of bacteria. F-actin was visualized by Texas Red-phalloidin (red), VAMP8 by YFP (green), Salmonella by anti-LPS antibody (blue). Bars represent 20 µm.

Figure 1 (.jpg)

Supplemental Figure 2: The SopB^C460S mutant strain is deficient in VAMP8 recruitment. MDCK cells were infected with the SopB^C460S mutant strain at an MOI of 10, VAMP8 was stained with rabbit anti-VAMP8 antibodies (green), and F-actin by Texas Red-phalloidin (red). Arrows point to sites of ruffle formation.

Figure 2 (.jpg)

Supplemental Figure 3: Expression levels of recombinant proteins. The expression level of GST-p40-PX, GST-VAMP8^1-79 and His-YFP-VAMP8 were comparable in E. coli lysate (left panel). The expression level of YFP, YFP-VAMP8 and YFP-VAMP8^1-79 were also similar in HeLa cell lysates (right panel).

Figure 3 (.jpg)