Fig. 4: Metabolic profiles of Mtb-infected AMs and KCs.
AMs and KCs from C57BL/6 mice were isolated and infected with Mtb H37Rv. After 72 h, cell lysates were analyzed using LC-MS. A representative score plot of the partial least squares discriminant analysis (PLS-DA) was generated using MetaboAnalyst. PLS-DA models were validated using R2 and Q2 based on LOOCV (leave one out cross-validation); the four-component model was selected as the optimized model with R2 = 0.95 and Q2 = 0.58. The significance of the model was demonstrated by a permutation test with 100 testing iterations using a separation distance of p < 0.01. a AMs control (red); AMs Mtb H37Rv (green). b KCs control (red); KCs Mtb H37Rv (green). c AMs Mtb H37Rv (red); KCs Mtb H37Rv (green). d Representation of 25 metabolites with VIP (variable importance of projection) scores greater than 1.0 based on PLS-DA considered significant. On the extreme right, red and green indicates high and low levels of metabolites, respectively. e Quantitative metabolite sets enrichment overview using metabolite set enrichment analysis (MSEA) with the fold change showing the metabolic pathways of 25 metabolites selected based on their VIP scores greater than 1. The arrow indicates upregulated metabolites and pathways in Mtb H37Rv-infected KCs. f KCs from C57BL/6 mice were isolated and infected with Mtb H37Rv at an MOI of 1:2.5 (1 AM and 2.5 Mtb). At 2 h post-infection, nor-NOHA were added at indicated concentrations. Intracellular CFUs were demonstrated 5 days post-infection. Statistical analysis was performed with paired two-tailed t test and presented as mean ± SD. Data shown are representative of five independent experiments.
