Extended Data Fig. 9: Metabolic effects on surface BTN3A expression.
From: CRISPR screens decode cancer cell pathways that trigger γδ T cell detection
(a) Surface BTN3A MFI in Daudi-Cas9 KOs cultured in different pyruvate concentrations for 3 days in RPMI (no glucose, no pyruvate). Normalized to cells grown without pyruvate (0 mM). (b-d, f) Surface BTN3A MFI in Daudi-Cas9 cells treated for 72 h with (b) an mTOR inhibitor (rapamycin), an ISR inhibitor (ISRIB), ISR agonists (guanabenz, Sal003, salubrinal, raphin1, sephin1), and DMSO (vehicle) (KO cells); (c) metformin (WT cells); (d) A-769662 compared to equivalent amounts of DMSO (vehicle) (WT cells); or (f) the shown compounds (KO cells). (e) Surface BTN3A MFI in WT Daudi-Cas9 cells co-treated with AICAR and increasing amounts of Compound C (AMPK inhibitor) or DMSO (vehicle). (a) n = 4 per condition (n = 3, TIMMDC1 (#2) at 0 mM), data combined from two independent experiments, each individually normalized. (b) n = 6 per condition (except n = 5 for AAVS1 (#5) with guanabenz and for PPAT (#1) with salubrinal), data combined from two independent experiments, each individually normalized to DMSO (vehicle)-treated cells. (c) n = 8 per condition, data combined from two independent experiments. One-way ANOVA comparison to cells that received no treatment with Dunnett’s multiple comparisons test. (d) n = 3 per condition, representative data from one of two independent experiments. (e) n = 3 per conditions, representative data from one of two independent experiments. Two-tailed unpaired Student’s t test with Bonferroni correction. (f) n = 3 per condition (n = 2 for AMPKα1 (#1) treated with DMSO), representative data from one of two independent experiments. One-way ANOVA comparison to AAVS1 (#5) KO cells with Dunnett’s multiple comparisons test. (a-f) Mean ± SD. p < 0.0001 (****), p < 0.001 (***), p < 0.01 (**), p < 0.05 (*), p > 0.05 (N.S.).
