Fig. 3: MFN2 is required for the induction of Hif1α and Ldha in macrophages.

a PLS-DA score plots for metabolites in Mfn2 WT and Mfn2 CKO BMDMs between uninfected and after Mtb infection (MOI 5) based on GC-TOF/MS (VIP > 0.7, p value <.05). Each group designate with a different color (red: untreated Mfn2 WT, orange: Mtb-infected Mfn2 WT, green: untreated Mfn2 CKO and blue: Mtb-infected Mfn2 CKO). b The bar plots showed relative concentration of identified organic acids that was calculated by relative peak area from GC-TOF/MS analysis. c Heatmap analysis shows the differentially expressed genes in Mfn2 WT and Mfn2 CKO BMDMs before and after infection of Mtb for 18 h. The z-scores were calculated for each gene row using the average gene abundances of biological replicates. d Bar graph shows the abundance of genes Ldha, Ldhb, and Hif1a between Mfn2 WT and Mfn2 CKO BMDMs (n = 4). e, f qPCR analysis of Ldha (e) and Hif1a (f) mRNA expression in Mtb (MOI 5) and LM (MOI 5) infected Mfn2 WT or Mfn2 CKO BMDMs (n = 6). g, h Extracellular acidification profile (g) and representative glycolysis capacity parameter (h, n = 5) in Mfn2 WT and Mfn2 CKO PMs infected with Mtb (MOI 5) for 18 h. i Extracellular lactate level in Mfn2 WT and Mfn2 CKO BMDMs after Mtb (MOI 5) infection for 18 h. Data are representative or pooled from three independent experiments and are presented as mean ± SEM. The data were analyzed by the multiple t test with Holm–Sidak correction (b) or by two-tailed Student’s t test (d–f, h, i). Un, uninfected.