Supplementary Figure 3 from Chromatin Helicase CHD6 Establishes Proinflammatory Enhancers and Is a Synthetic Lethal Target in FH-Deficient Renal Cell Carcinoma

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Supplementary Figure 3. CHD6 is essential for FH-deficient RCC cells, related to Figure 4. A-B. MTT analysis of FH-WT PRCC (A) or canonical ccRCC (B) cells with or without CHD6-KD. C. Competition-based assay to measure the effect of CHD4 shRNA on the growth of UOK-262 cells (n = 3 per time point). CHD4-KD cells were identified by coexpression of green fluorescent protein (GFP) (LMN vector). The percentage of GFP+ cells was thus tracked over 12 days and normalized to GFP percentage on day 2. D. Quantified BIL signals of orthotopic (FH-intact Caki-2) renal tumors with or without CHD6 ablation. E. Serum VEGF concentrations in treated BALC/c nude mice from indicated groups were compared. F. Ki-67 (IHC) and TUNEL (IF) staining from xenografts derived from FH-KD and FH/CHD6-KD Caki-2 cells, and the quantitative results are shown in the right panel. Scale bar, 20 μm. G. Kaplan-Meier survival curve analysis of mice from Figure 4F. H. Schematic graph showing the zebrafish tumour xenograft model construction at each time point. I. Quantified tumour sizes derived from indicated cells implanted in the zebrafish embryos. J. Quantitation of organoid sizes from human FH-WT or FM-RCC samples with or without CHD6 depletion. P values were calculated using 2-way ANOVA followed by Tukey’s multiple comparisons tests (A, B), 2-tailed Student’s t-test (C-F, I-J), and log-rank test (G). *p < 0.05, **p < 0.01, and ***p < 0.001. ns, no significance.

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