Fig. 5: VHL reconstitution restores BCAA functioning in ccRCC cells.

a Heatmap showing the enriched pathways in the indicated comparisons obtained through GSEA analysis of proteomics data generated from cells grown in RPMI. b Volcano plot showing the differential expression of proteins that belong to KEGG “Valine leucine and isoleucine degradation” signature in 786-O + VHL vs. 786-O + EV and 786-M1A + VHL vs. 786-M1A + EV from proteomics data obtained culturing cells in RPMI. FC fold-change, red = upregulated proteins, blue = downregulated proteins. c Ratio of the intracellular abundance of C3-carnitines and C5-carnitines in cells expressing VHL compared to EV cultured in RPMI. Data were normalized to total ion count and represent the mean of 3 independent experiments (N = 3) ± S.E.M. p-values were calculated using two-tailed one-sample t-test against the theoretical mean of 1 (786-O + EV = 1 vs. 786-O + VHL and 786-M1A + EV = 1 vs. 786-M1A + VHL). d Proportion of total pool of the intracellular ketoisocaproic acid (KIC) and C5-carnitine derived from ¹³C leucine in renal cells cultured in RPMI. Data represents the mean of 5 independent cultures +SD. e mRNA levels of SLC7A5 in the indicated cell lines grown in RPMI measured through qPCR. TBP was used as endogenous control. Values represent relative quantification (RQ) ± error calculated using Expression suite software (Applied Biosystem) calculated using SD algorithm. p-value was calculated through Expression suite software. N = 3 independent experiments.