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. Author manuscript; available in PMC: 2019 Jul 1.
Published in final edited form as: Sci Signal. 2019 Jan 1;12(562):eaau9048. doi: 10.1126/scisignal.aau9048

Figure 3. GPR35 controls macrophage and intestinal epithelial cell metabolism.

Figure 3

A, Glucose uptake in M0, M1 and M2 BMDMs in the absence or presence of ouabain (1 mM). Glucose uptake expressed as intracellular 2-deoxy-D-glucose-6-phosphate (2DG6P) concentration detected per cell; n=3 mice/group. B, Lactate production measured in M1 and M2 BMDMs; n=3 mice/group. C, ECAR measured during sequential injection of oligomycin (O) and 2-deoxyglucose (2-DG) in Caco2 cells transfected with control (ctrl) siRNA (red) or GPR35 siRNA (blue). Bar chart shows basal ECAR; n=3 mice/group. D, Cellular respiration measured as basal and maximal oxygen consumption rate (OCR) during sequential treatment with oligomycin (O), FCCP (F) and antimycin/rotenone (A/R) in M0 and M2 BMDMs. Bar chart indicates basal and maximal OCR after FCCP injection; n=3 mice/group. E, Basal and maximal OCR measured during sequential treatment with oligomycin (O), FCCP (F) and antimycin/rotenone (A/R) in Caco2 cells transfected with either control (ctrl) siRNA (red) or GPR35 siRNA (blue). Bar chart indicates basal and maximal OCR after FCCP stimulation; n=3 independent experiments/group. F, Mitochondrial membrane potential (ΔΨm) in M0, M1 and M2 BMDMs in the absence or presence of 1 mM ouabain. Depolarisation with 1 μM FCCP used as a control. ΔΨm expressed as relative fluorescent units (RFU) at 590 nm; n=6 mice/group. G, Glucose uptake in human iPSC derived M0 macrophages in the absence or presence of ouabain (1 mM); n=3 independent experiments. H, Basal ECAR measured in human iPSC derived M0 macrophages; n=8 sets of cells/group. I, Lactate production in human iPSC derived M0 macrophages; n=3-4 sets of cells/group. J, Basal and maximal OCR during sequential treatment with oligomycin (O), FCCP (F) and antimycin/rotenone (A/R) in iPSC-derived human macrophages homozygous for the GPR35108T (red) or GPR35T108M (black) alleles. Bar chart indicates basal and maximal OCR after FCCP stimulation; n=8 sets of cells/group. All data represented as mean ± s.e.m. Statistical significance was calculated by two-tailed Student’s t test using individual data points, * (p<0.05), ** (p<0.01).