pknF Mtb mutant infection compared to Mtb infection does not induce increase phagosomal membrane rupture and/or cytosolic access in BMDM.

BMDMs were infected with CDC1551 Mtb and ΔpknF mutant at an MOI of 10 for 4h. At 6 hpi, cells were fixed, permeabilized and immunostained for TAX1BP1. Colocalization between TAX1BP1 and different Mtb strains (Mtb and ΔpknF mutant) was (A) imaged using LSM 980 Laser scanning confocal microscope, Scale bar 5μm and (B) quantified by determining the Pearson’s correlation coefficient (r) with ImageJ JaCoP plug-in in 36 randomly selected fields of view from each Mtb strain that included 4 to 5 cells per field in three independent experiments (A value of -1 indicates perfect exclusion, zero represents random localization, while +1 indicates perfect correlation). BMDMs derived from wild type (WT) and Asc^-/- mice were either left uninfected (UI) or infected with Mtb and ΔpknF mutant at an MOI of 10 for 4h. The culture supernatants were harvested at 20 hpi and analyzed for (C) IFN-β levels by ELISA. Data are representative of three independent experiments. Error bars represent mean ± SEM; **, p<0.01, ***, p<0.001, ns (non-significant).

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