Fig. 5: GATD3A deficiency reduces TCA cycle flux, altering the metabolic pattern of FLSs.

a Heatmap of label-free proteomics assay for Nor-FLSs infected with ShNC or ShGATD3A lentivirus. b Number of proteins involved in biological processes was analysed through GO enrichment analysis based on proteomic assay. c KEGG pathway enrichment analysis showed the number of proteins interacting with GATD3A within these pathways based on IP/MS. OPLS-DA analysis (d) and KEGG pathway enrichment analysis (e) of the untargeted metabolomics study for Nor-FLSs infected with ShNC or ShGATD3A lentivirus. f Heatmap of significantly different metabolites in the TCA cycle, glutaminolysis and glycolysis based on the untargeted metabolomics study for Nor-FLSs infected with ShNC or ShGATD3A lentivirus. OCR (n  =  5 for glucose supplementation group, and n = 4 for other two groups) (g) and PER (n  =  6) (h) assays of Nor-FLSs treated with glucose or glutamine, and glucose and glutamine deprivation. i Diagram of ¹³C isotopomer patterns with [¹³C₅] glutamine and [U-¹³C₆] glucose as tracer. j Absolute isotopologue distribution of [¹³C₅] glutamine-derived glutaminolysis and TCA cycle metabolites including glutamate, α-KG, succinate, fumarate, malate, oxaloacetate, citrate and aspartate. n  =  4 independent biological replicates. The incorporation of ¹³C atoms was denoted as m + n (number of ¹³C atoms). k Absolute isotopologue distribution of [U-¹³C₆] glucose-derived glycolysis metabolites including pyruvate and lactate. n  =  4 independent biological replicates. The incorporation of ¹³C atoms was denoted as m + n (number of ¹³C atoms). l Cellular malate and oxaloacetate content were determined in Nor-FLSs infected with ShNC or ShGATD3A lentivirus. n  =  5 independent biological replicates. The glucose uptake rate (n  =  5 independent biological replicates) (m) and the cellular lactate content (n  =  4 independent biological replicates) (n) in Nor-FLSs infected with ShNC or ShGATD3A lentivirus. All data were presented as the means ± SD. P values were determined by two-tailed unpaired Student’s t-test or one-way ANOVA with Tukey’s multiple-comparisons (g, h). P < 0.05 was considered statistically significant. Source data are provided as a Source Data file.