Figure 4.
Characterization of Mtb Infection in iMACs and Anti-TB Drug Responsiveness
(A) Confocal micrographs showing the percentage of iMACs infected with GFP-labeled Mtb at various initial MOIs in the presence and absence of the anti-TB drugs INH (10 μM) or RIF (10 μM) after 5 days of infection. Infected cells harbor H37Rv-GFP bacteria (green) and are stained with DAPI (blue).
(B) Percentage of Mtb-infected iMACs obtained at various MOIs after 5 days infection, in the presence or absence of anti-TB drugs (INH or RIF). Data represent mean ± SD of three independent experiments.
(C) The correlation between the imaging method and the CFU counting method for iMACs infected with H37Rv-GFP strain. Average ± SD values are shown; n = 4 wells for the ratio of infected cells, n = 2 wells for the CFUs.
(D and E) Quantification by ELISA of secreted cytokines in culture medium of iMACs 5 days after Mtb infection (MOI 5). TNF-α (D) and IL-6 (E) concentrations are shown in pg/mL. Eight-fold diluted culture medium were used.
(F–H) Heatmap of differentially expressed genes (p < 0.05) of hMDMs on day 5 after Mtb infection (MOI 5) detected by RNA-seq, respectively (F). Annotated in GO (GO terms; innate immune and inflammatory responses genes [GO: 0045087, 0006954]). Color scale indicates log2 fold change. qRT-PCR analysis represents relative fold change (RFC) of genes expression after Mtb infection in hMDMs (G) or iMACs (H). p < 0.005 (n = 3).