Extended Data Fig. 1: LRRK2 is required for efficient itaconate delivery to the Salmonella-containing vacuole.
(a and b) LRRK2 is required for efficient itaconate delivery to the Salmonella-containing vacuole. Parental (control) and Lrrk2-/- Raw264.7 cells were infected with S. Typhi (MOI = 6) encoding an eGFP-based itaconate biosensor and the number of cells expressing eGFP was determined 20 hours after infection. Each square and circle represents the mean of an individual experiment experiments in which at least 200 infected cells were examined (b). The p value (unpaired two-tailed Student’s t test) of the indicated comparison is shown. Infected cells were fixed, stained with DAPI (blue) to visualize nuclei, and stained with an anti-Salmonella LPS antibody along with Alexa 594-conjugated anti-rabbit antibody (red) to visualize all bacteria. Representative fields of infected cells are shown (a) (scale bar = 5 µm). (c-g) Absence of LRRK2 does not influence the uptake of Salmonella into phagocytic cells. Raw264.7 or DC2.4 parental (control) and Lrrk2-/- cells, as well as bone marrow-derived macrophages (BMDM) derived from C57BL/6 and Lrrk2-/- mice were infected with either wild-type S. Typhi (MOI = 6) or a S. Typhimurium ∆gtgE ∆sopD2 mutant strain (MOI = 3) (as indicated) and the number of CFU was determined 1 hr after infection. Each square or circle represents the CFU in an independent measurement. The mean ± SD and p values (unpaired two-tailed Student’s t test) of the indicated comparisons are shown (n = 6 for each category).
