Fig. 1: Generation and validation of pdCas9a-SAM construct in Eµ-Myc lymphoma cell lines. | Nature Communications

Fig. 1: Generation and validation of pdCas9a-SAM construct in Eµ-Myc lymphoma cell lines.

From: Generation of a CRISPR activation mouse that enables modelling of aggressive lymphoma and interrogation of venetoclax resistance

Fig. 1

a Schematic representation of the pdCas9a-SAM lentiviral construct. b Western blot analysis for BCL-2 in manipulated Eµ-Myc lymphoma cell lines. EMRK-1184 or MRE-721 Eµ-Myc lymphoma-derived cell lines were transduced with pdCas9a-SAM only or pdCas9a-SAM plus Bcl-2 sgRNAs. Cell lysates were harvested from transduced cell lines, before or after puromycin-selection and expression of the indicated proteins was examined by Western blotting. Probing for the heat shock protein 70 (HSP70) served as a loading control. 2 independent experiments were performed showing similar results. c, d Viability of Eµ-Myc lymphoma cell lines transduced with pdCas9a-SAM only or pdCas9a-SAM plus Bcl-2 sgRNAs. Cells were treated with the MCL-1 inhibitor S63845 or the BCL-2 inhibitor venetoclax at the indicated drug concentrations. Cell viability was determined at 24 h by propidium iodide (PI) staining and subsequent flow cytometric analysis. Data are represented as mean ± SD, n = 3 independent experiments. The statistical significance was determined using two-way ANOVA test with EMRK-1184 pdCas9a-SAM or MRE-721 pdCas9a-SAM cells serving as the control. Exact P values are provided in the Source Data file. Overall P values are shown as ns = no significant difference, **P < 0.01. Source data are provided as a Source Data file.

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