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. 2021 Nov 10;10:e73984. doi: 10.7554/eLife.73984

Figure 2. Infection with Mycobacterium tuberculosis (Mtb) phthiocerol dimycocerosate (PDIM) or ESX-1 mutants elicits enhanced expression of an inflammatory transcriptional program.

(A) FPKM from RNAseq data (MOI 2:1) for representative genes from subcluster 1B. *p-value < 0.01 or #p-value < 0.05 for the comparison of PDIM (16 hr) or ESX-1 (12 hr) mutant-infected with Rv-infected, unpaired two-tailed t-test. (B) C57BL/6J bone marrow-derived macrophages (BMDM) were infected with the indicated strains at an MOI of 10:1. RNA was harvested at the indicated timepoints, and expression of the indicated genes was profiled by qPCR relative to GAPDH control. (C–D) C57BL/6J BMDM were infected with the indicated Mtb strains at an MOI of 2:1. RNA was harvested 24 hr post-infection, and expression of the indicated genes relative to GAPDH control was profiled using qPCR. (B–D) Mean ± SD of four replicates. *p-value < 0.01, **p-value < 0.001, ***p-value < 0.0001 unpaired two-tailed t-test. RNAseq experiment (A) performed once, (B) one of two independent experiments, (C–D) one of three independent experiments.

Figure 2.

Figure 2—figure supplement 1. Enhanced expression is independent of BMDM mouse strain and specific to PDIM and ESX-1 mutants.

Figure 2—figure supplement 1.

(A, C) BALB/c (A) or C57BL/6J (C) bone marrow-derived macrophages (BMDM) were infected with the indicated Mycobacterium tuberculosis (Mtb) strains at an MOI of 2:1. RNA was harvested 24 hr post-infection, and expression of the indicated genes was quantified by qPCR relative to GAPDH control. (B) J774 cells were infected with the indicated Mtb strains at an MOI of 2:1. Cells were lysed at day 0 and day 3; lysates were plated for CFU. (C) C57BL/6J BMDM were infected with the indicated Mtb strains at an MOI of 2:1. (A–C) Mean ± SD for four replicates. ***p-value < 0.0001 unpaired two-tailed t-test.