Fig. 6: SOX6-mediated TXNIP expression causes Elesclomol sensitivity.
a Correlation of TXNIP and SOX6 expression in 18 EwS cell lines. The line indicates the linear regression of the data (two-tailed Pearson correlation test, P = 0.016, rpearson = 0.559). b Top: Representative Western blot analysis of TXNIP 96 h after SOX6 knockdown. GAPDH served as loading control. Bottom: Relative TXNIP expression (qRT-PCR). Horizontal bars represent means, n = 3 biologically independent experiments. P values determined via two-sided Mann–Whitney test (all P = 0.002). c Top: representative micrographs from EwS xenografts with/without SOX6 knockdown stained for TXNIP. Bottom: IRS for TXNIP (n indicates the number of biologically independent xenografts per condition: TC-32: shCtrl (Dox(–) n = 3, Dox(+) n = 4), shSOX6_2/3 (Dox(–) n = 11, Dox(+) n = 8, P ≤ 0.0001); RDES: shCtrl (Dox(–) n = 4, Dox(+) n = 6), shSOX6_2/3 (Dox(–) n = 7, Dox(+) n = 8, P = 0.007). Horizontal bars represent medians. P values determined via two-sided Mann–Whitney test. Scale bar = 20 µm. d Relative TXNIP expression (qRT-PCR) in TC-32 cells 96 h after TXNIP knockdown. Horizontal bars represent means, n = 5 biologically independent experiments, P = 0.008. P value determined via two-sided Mann–Whitney test. e Relative DCF-DA fluorescence in TC-32 cells after TXNIP knockdown. Horizontal bars represent means, n = 5 biologically independent experiments, P = 0.009. P value determined via two-sided independent one-sample t-test. f MitoSOX Red/MitoTracker Green ratio in TC-32 cells after TXNIP knockdown. Horizontal bars represent means, n = 4 biologically independent experiments, P = 0.012. P value determined via two-sided independent one-sample t-test. g Elesclomol IC50 upon TXNIP knockdown in TC-32 cells. Horizontal bars represent means, n = 4 biologically independent experiments, P = 0.029. P value determined via two-sided Mann–Whitney test. h Left: Relative mRNA expression in TC-32 EwS cells after 96 h of concomitant Dox-treatment (0.1 µg/ml) and Cumate-treatment (5 µg/ml). Horizontal bars represent means, n = 5 biologically independent experiments, P = 0.008. Right: Relative Elesclomol IC50 in TC-32 cells 72 h after Elesclomol-treatment and concomitant addition of Dox (0.1 µg/ml) and Cumate (5 µg/ml). Horizontal bars represent means, n = 5 biologically independent experiments, P = 0.008. P values determined via two-sided Mann–Whitney test. i Schematic illustration of the EWSR1-FLI1-mediated effect of SOX6 expression. All error bars represent SEM. ****P ≤ 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05. Source data are provided as a Source Data file.
