Figure 5.

ARNO Generation of Salmonella-Induced Macropinosomes

(A) Localization of ARNO during Salmonella invasion. HeLa cells expressing ^CFPARNO (ARNO) were infected for 15 min with fluorescently labeled WT or ΔE/E2 Salmonella (bacteria) as indicated (left). Scale bar 8 μm. Insets show magnified area.

(B) ARNO recruitment by Salmonella. To quantify ARNO recruitment, HeLa cells expressing either ^CFPARNO (ARNO), ^CFPARNO^R279C (R279C), ^CFPARNO^K336A (K336A), or ^CFPARNO^R279CK336A (R279C K336A) were infected with fluorescently labeled WT Salmonella, and the proportion of bacteria enriched with ARNO was determined microscopically. Asterisks indicate a significant difference from mock (p < 0.05, ANOVA; n = 3). Error bars represent ± SEM.

(C) Quantification of ^CFPARNO (ARNO) recruitment by WT, ΔinvG, ΔE/E2, and ΔsopB Salmonella strains was performed as in (B). See also Figure S3.

(D and E) Localization of ARNO, Arf1, and HSPC300 during Salmonella invasion. HeLa cells expressing ^CFPARNO and Arf1^RFP (Arf1) (D) or ^CFPHSPC300 (HSPC300) and Arf1^RFP (E) were infected as (A).