Figure S1.
Funtional analysis of SETD1B loss in DLBCL cell lines. (A) Bar graphs displaying the validation of the CRISPR-cas9 screen hits with individual sgRNA by cell competition assay. OCI-LY19 cells were partially transduced with sgRNA targeting top candidate genes from the genome-wide screening. sgRNAs co-expressing tRFP (turbo-red fluorescent protein) and the percentages of tRFP-positive cells were measured by flow cytometry. Cells were treated with DMSO or 100 nmol Venetoclax for 3 days and recovered for 3–5 days and the treatment and recovery cycle was repeated once. The percentages of tRFP-positive cells after treatment were normalized to before treatment as FC to indicate the Venetoclax resistance of cells transduced with sgRNAs targeting these candidate genes. Results were represented as mean ± SD (n = 3). (B) IC50 curves of MCL-1 inhibitor, S63845, in OCI-LY19 cells carrying only Cas9, or Cas9; sgLacZ, or Cas9; sgSETD1B single-cell clones from two independent sgRNAs. Values represent the average of three independent dose-titration curves (n = 3). (C) Representative immunoblotting of OCI-LY19; sgLacZ and OCI-LY19; sgSETD1B clones were probed with SETD1B and BCL2 family proteins. β-Actin was used as a loading control. Data representative of at least two independent experiments are shown. MW, molecular weight in kD. (D) Annexin V analysis of OCI-LY19; sgLacZ and OCI-LY19; sgSETD1B clones treated with DMSO or 2.5 μM Venetoclax. Results were represented as mean ± SD (n = 3). (E) IC50 curves of Venetoclax in OCI-LY8 cells carrying only Cas9, or Cas9; sgLacZ, or Cas9; sgSETD1B single-cell clones from two independent sgRNAs. Values represent the average of three independent dose-titration curves (n = 3). (F) Annexin V analysis of OCI-LY8; sgSETD1B and OCI-LY8; sgLacZ treated with DMSO or 2.5 μM of Venetoclax. Values represent means and SD (n = 3). (G) IC50 curves of SU-DHL-10 isogenic cell lines with (sgSETD1B clone 1 and clone 2) or without (sgLacZ) SETD1B mutations after 72 h of Venetoclax treatment. Values represent means and SD (n = 3). (H) IC50 curves of OCI-LY8 isogenic cell lines with SETD1B (sgSETD1B), KMT2D (sgKMT2D), or with SETD1B and KMT2D mutations (sgSETD1B;KMT2D) after 72 h of Venetoclax treatment. Values represent means and SD (n = 3). Source data are available for this figure: SourceData FS1.

Funtional analysis of SETD1B loss in DLBCL cell lines. (A) Bar graphs displaying the validation of the CRISPR-cas9 screen hits with individual sgRNA by cell competition assay. OCI-LY19 cells were partially transduced with sgRNA targeting top candidate genes from the genome-wide screening. sgRNAs co-expressing tRFP (turbo-red fluorescent protein) and the percentages of tRFP-positive cells were measured by flow cytometry. Cells were treated with DMSO or 100 nmol Venetoclax for 3 days and recovered for 3–5 days and the treatment and recovery cycle was repeated once. The percentages of tRFP-positive cells after treatment were normalized to before treatment as FC to indicate the Venetoclax resistance of cells transduced with sgRNAs targeting these candidate genes. Results were represented as mean ± SD (n = 3). (B) IC50 curves of MCL-1 inhibitor, S63845, in OCI-LY19 cells carrying only Cas9, or Cas9; sgLacZ, or Cas9; sgSETD1B single-cell clones from two independent sgRNAs. Values represent the average of three independent dose-titration curves (n = 3). (C) Representative immunoblotting of OCI-LY19; sgLacZ and OCI-LY19; sgSETD1B clones were probed with SETD1B and BCL2 family proteins. β-Actin was used as a loading control. Data representative of at least two independent experiments are shown. MW, molecular weight in kD. (D) Annexin V analysis of OCI-LY19; sgLacZ and OCI-LY19; sgSETD1B clones treated with DMSO or 2.5 μM Venetoclax. Results were represented as mean ± SD (n = 3). (E) IC50 curves of Venetoclax in OCI-LY8 cells carrying only Cas9, or Cas9; sgLacZ, or Cas9; sgSETD1B single-cell clones from two independent sgRNAs. Values represent the average of three independent dose-titration curves (n = 3). (F) Annexin V analysis of OCI-LY8; sgSETD1B and OCI-LY8; sgLacZ treated with DMSO or 2.5 μM of Venetoclax. Values represent means and SD (n = 3). (G) IC50 curves of SU-DHL-10 isogenic cell lines with (sgSETD1B clone 1 and clone 2) or without (sgLacZ) SETD1B mutations after 72 h of Venetoclax treatment. Values represent means and SD (n = 3). (H) IC50 curves of OCI-LY8 isogenic cell lines with SETD1B (sgSETD1B), KMT2D (sgKMT2D), or with SETD1B and KMT2D mutations (sgSETD1B;KMT2D) after 72 h of Venetoclax treatment. Values represent means and SD (n = 3). Source data are available for this figure: SourceData FS1.

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