Extended Data Fig. 7: Rv1272c induced linoleic acid promotes CTLA-4 surface trafficking on Tregs to inhibit cytosol ROS and facilitate the survival of Mtb in macrophages.
(a) Mtb-infected BMDMs (MOI = 5) were pretreated with indicated inhibitors for 24 h and then co-cultured with Tregs treated with isotype control IgG or anti-CTLA-4 blocking antibody (40 µg/mL) for 1 day, intracellular survival of the indicated Mtb strains in BMDMs infected for 2 days were determined using c.f.u. assay. (b) After 1 day of infection with indicated Mtb strains (MOI = 5), BMDMs were co-incubated with CD4+ ESAT6-specific Treg cells treated with or without linoleic acid for 24 h, MFI of membrane CTLA-4 levels in CD4+ Tregs were measured by FCM. (c) After 1 day of infection with indicated Mtb strains (MOI = 5), BMDMs were co-incubated with CD4+ ESAT6-specific Treg cells treated with or without linoleic acid for 24 h. MFI of ROS levels in BMDMs were measured by FCM. Data in a-e represent one experiment with at least three independent biological replicates (a, n = 4; b-c, n = 3); mean ± s.e.m. Two-tailed unpaired Student’s t-tests (b) and Two-way ANOVA with Tukey’s multiple comparisons test (a, c) were used for statistical analyses.
