Extended Data Fig. 2: Proline depletion upon P5CS/Aldh18a1 downregulation.
A) Relative abundance of intracellular PRO in WT mouse kidney epithelial cells from two different thaws (WT-1 and WT-2) and two P5CS KO subclones (C2 and C9) infected either with empty vector ( + Vec) or reconstituted with P5CS ( + P5CS). Cells were grown for 24 hours in proline-free high-glucose DMEM medium with 0.65 mM GLN and 2.5%FBS, and metabolite levels normalized by total protein (n = 3) Statistical significance was determined using an unpaired, Two-tailed T-Test. Data are presented as mean values ± SD. B) Metabolomic profiling of amino acid consumption and secretion. This panel of plots illustrates the relative consumption and secretion rates of various amino acids by control and P5CS KD cells, under low-glutamine conditions. The lower bar graphs show the consumption rates of amino acids from the media, while the upper ones depict the secretion rates of amino acids into the media. Data are represented as mean ± SD of three independent experiments. The statistical analysis was performed using two-way ANOVA.
