Fig. 1.

Metabolomic profiles of ICL-deficient Mtb when cultured in media containing different carbon sources. Intrabacterial pool sizes and isotopic labeling of metabolic intermediates of glycolysis/gluconeogenesis, TCA cycle, glyoxylate shunt, and methylcitrate cycle in wild-type (Erdman), ICL1/2-deficient (KO), and ICL-reconstituted (COM) Mtb strains incubated in [U-¹³C]-containing media for 24 h. Total bar heights indicate the intrabacterial concentration, whereas the colored area of each bar denotes the extent of ¹³C labeling achieved following transfer to [U-¹³C] dextrose (green) (A) and [U-¹³C] propionate (purple)-containing media (B) under the condition indicated. Schematic diagram when transferring to [U-¹³C] acetate (red)-containing media is shown in Fig. S2. ICL encodes both isocitrate lyase (gray dotted line) and methylisocitrate lyase (blue dotted line). All values are average of three independent experiments, each of which consisted in biological triplicates ± SE. Intrabacterial pool sizes of metabolites analyzed were described as nmol/mg except for 2 methyl cis-aconitate, whose relative abundance is instead reported as ion counts per milligram, due to lack of a commercially available standard. ***P < 0.001; **0.001 < P < 0.01; *0.01 < P < 0.05; and ns, not significant by ANOVA. αKG, α-ketoglutarate; 2MC, 2 methylcitrate; 2M-ACO, 2 methyl cis-aconitate; 2MIC, 2 methylisocitrate; Asp, aspartate; cis-ACN, cis-aconitate; FBP, fructose 1,6-bisphosphate; FUM, fumarate; GLO, glyoxylate; Hex-P, hexose phosphate; IC, ion counts; MAL, malate; Pent-P, pentose 5-phosphate; PEP, phosphoenolpyruvate; PYR, pyruvate; SH7P, sedoheptulose 7-phosphate; Triose-P, triose 3-phosphate (glyceraldehyde 3-phosphate and dihydroxyacetone phosphate).