Fig. 5

SCF-c-Kit signalling drives neutrophil metabolic adaptations. a Tumour weight and b spleen weight following administration of anti-c-Kit antibody (α c-Kit) (50 µg) or an isotype control (50 µg). Data were analysed by one-way ANOVA with Tukey’s multiple comparisons displayed on graph. Tumour n = 10, spleen control n = 15, isotype and anti-c-Kit n = 10, from two independent experiments. c Percentage Ly-6G positive leukocytes in blood. Data were analysed by one-way ANOVA with Dunnett’s multiple comparisons represented on graph, isotype and anti c-Kit n = 5, naive n = 10. d Representative neutrophil mitochondrial stress test (oligomycin (Oligo, 1.26 µM), Carbonyl cyanide-4(trifluoromethoxy)phenylhydrazone (FCCP, 660 nM), Rotenone (R, 100 nM) and antimycin A (AA, 1 µM) displaying oxygen consumption rates (OCR) with indicated treatments. e ATP synthase-dependent OCR displaying the proportion which is sensitive to inhibition of fatty acid oxidation (FAO) with etomoxir (100 µM). Data (n = 3) from two independent experiments were analysed by two-way ANOVA with Tukey’s multiple comparisons indicated. f Representative OCR traces following stimulation with phorbol 12-myristate 13-acetate (PMA, 1 µg/ml) (arrow) in the presence or absence of 2-deoxyglucose (2DG, 100 mM) (line). g Area under the curve (AUC) of neutrophil OCR from data in (f), data shows relative OCR in the presence of 2-DG compared to PMA alone. Data (n = 3) from two independent experiments. Data were analysed by one-way ANOVA with Dunnett’s multiple comparisons indicated. h Tumour weights and i spleen weights following injection of 4T1 with CRISPR silencing of kitl (51-1) or a non-silenced control (50-1). Data (n = 10) from two independent experiments, data were analysed by unpaired t test. j Percentage of Ly-6G positive leukocytes in blood and k percentage of neutrophils which are c-Kit⁺. Data (n = 10) from two independent experiments, data were analysed by unpaired t test. l ATP synthase-dependent OCR displaying the proportion which is sensitive to inhibition of FAO with etomoxir (100 µM). Data (n = 10) from two independent experiments, data were analysed by two-way ANOVA with Sidak’s multiple comparisons indicated. m Relative AUC of neutrophil OCR following PMA stimulation in the presence of 2-DG compared to PMA alone. Data (n = 10) from two independent experiments. Data were analysed by by unpaired t test. p Values *<0.05, **<0.01, ***<0.001. All error bars display mean ± SEM