Figure 5. Pyruvate carboxylation is a vulnerable adaptation to Sdhb loss.

(a) Aspartate abundance in PCX-silenced cells cultured for 24h with U-¹³C-glucose. Data are presented as mean±s.e.m of n=9 wells pooled from three independent experiments. (b) Relative abundance of ¹³C₁-aspartate in PCX-silenced cells incubated with ¹³C-bicarbonate for 10 min. Data are presented as mean±s.e.m of n=3 wells of one representative experiment, independently replicated twice. (c) Data are presented as mean±s.e.m of n=12 wells pooled from three independent experiments.. (d) PCX expression in H-Ras^V12-transformed cells infected with lentiviruses expressing either a shNTC or shPcx-1 sequence prior to injection into nude mice. Data are presented as mean±s.e.m (n=3 wells) of one experiment, performed once. (e-g) In vivo growth of cells described in (d) xenografted in athymic nude mice. The % of tumour-free mice over time (e) and the tumour volumes of each xenografted mouse (f, g) are presented (n=8 Sdhb^fl/fl-shNTC, n=8 Sdhb^fl/fl-shPcx1, n=9 Sdhb^Δ/Δ-shNTC, n=9 Sdhb^Δ/Δ-shPcx1). The Log-rank (Mantel-Cox) test was used to calculate the statistical significance between curves in (e). A statistical permutation test was used to compare the statistical significance between curves of the selected genotypes in (f), as described in Methods. Data derive from one experiment, performed once. Raw data of independently repeated experiments are provided in Supplementary Table 3.