Figure S3 from Lactate Utilization Enables Metabolic Escape to Confer Resistance to BET Inhibition in Acute Myeloid Leukemia

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Figure S3: CRISPR technology disrupts target genes. Expressions of (A) BRD4, (B, C) LDHB, and (C) MCT1 were disrupted in the indicated cell lines by CRISPR. The percent indel efficiency of the genetic disruption was quantified by TIDE analysis of Sanger sequencing relative to scramble control after 72 hr. (B) Cells were treated with BETi (0.15 uM) for 48 hr and viability quantified by a fluorescence plate reader. and Each point represents the mean value obtained from cell lines from an individual experiment (n=3). (D) Cells were transfected with siRNA targeting MCT4 or a scramble control. Subsequently, cells were treated with BETi for 48 hr and MCT1 and MCT4 protein levels quantified by flow cytometry and viability quantified using a fluorescence plate reader. Representative histograms for MCT1 and MCT4 protein are provided. (A) Paired t test, (B) one- or (D) two-way ANOVA with Tukey multiple comparisons test (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001, ns = not significant).

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