Supplementary Figure 1 from Chromatin Helicase CHD6 Establishes Proinflammatory Enhancers and Is a Synthetic Lethal Target in FH-Deficient Renal Cell Carcinoma

Supplementary Figure 1. Library and cells used for in vivo epigenetic CRISPR screen, related to Figure 1. A. Schematic diagram of FH mutations and related distributions in the indicated protein domains. B. Intracellular fumarate levels were measured in a panel of indicated RCC cell lines. C. Lorenz curve showing the distribution of sgRNAs in the epigenetic-focused library. D. Workflow showing the generation of the UOK- or ACHN-clones without Cas9 for evaluating the distribution of guides that persist upon tumour formation from the tumour initiating cells (TICs), and the UOK- or ACHN-clones with Cas9 were used for further in vivo screens. E. Western blot showing superior Cas9 expressions in UOK-Cas9-Clone 5 and ACHN-Clone 4. F. Cell viability of UOK-Cas9-Clone 5 and ACHN-Cas9-Clone 4 transfected with sgRNAs targeting the essential gene CCND1. G. Representative tumor graph and growth curve of sgRNA library transduced UOK-Cas9-Clone 5 & ACHN-Cas9-Clone 4 subcutaneously injected into BABL/c nude mice. H. Percentage of maintained sgRNAs in mice injected with UOK-Clone 5 library (without Cas9) and ACHN-clone 4 library (without Cas9) cells. I. Volcano plot revealing the targets with altered sgRNA frequencies with a cutoff of p < 0.01 and log2 fold change (log2FC) > 2. J. MTT analysis showing the effects of siRNA KD of 9 CHD family genes on FH-deficient cells, respectively. The quantitative results shown are representative of 5 experiments. P values were calculated using a two-tailed unpaired t-test (F, J). *p < 0.05, **p < 0.01, and ***p < 0.001.