Fig. 2

Comparative analysis of metabolite and nucleotide alterations in Tb PF CDA-like-RNAi and OE cell lines. (a) Relative expression of key metabolites altered in the Tb PF CDA-like-RNAi line after 6 days of DOX induction compared to the parental Tb PF line, measured using various mass spectrometry techniques. Data are presented as mean ± SD from 4 independent biological replicates. Statistical significance was determined using Welch’s t-test (*p < 0.05, **p < 0.01, ***p < 0.001). (b) Relative abundance of different lipid classes in the TbCDA-like-RNAi line compared to the parental Tb PF line. Data are presented as mean ± SD from 4 independent biological replicates. Boxes represent interquartile ranges and whiskers denote maximum and minimum values. (c) Levels of rNTPs in Tb PF, Tb PF CDA-like-RNAi and Tb PF CDA-like-OE lines, measured by anion exchange chromatography after 2 days of DOX induction. (d) Pyrimidine dNTP levels measured by the polymerase-based assay in Tb PF CDA-like-RNAi and Tb PF CDA-like-OE lines, 4 and 2 days post-induction, respectively. Panels c and d present data as mean concentrations with error bars indicating ± SD from three independent experiments with triplicate technical replicates. Asterisks indicate significant differences calculated by ANOVA. **p < 0.01; ***p < 0.001. The statistical significance analyses were performed by comparing the RNAi + DOX and OE + DOX cell lines with the Tb PF cell line.