Extended Data Fig. 2: PD-1 inactivation enables oncogene-enforced glycolysis in vivo.
a, Schematic outline of metabolic imaging analysis experiment. b, Exemplary splenic T1-weighted sequence of an acutely induced ITK-SYKCD4-creERT2;Pdcd1−/− animals with indicated ROI used for 7T-MRI [1-13C]pyruvate and [1-13C]lactate measurements. ROI, region of interest. c, Percentage of viable lymphocytes in the peripheral blood of anti-PD-L1 or isotype control antibodies treated ITK-SYKCD4-creERT2 mice. All animals received a single dose of tamoxifen on day zero. Cross indicates death of the animal (n = 3 mice per genotype). d, Schematic outline of metabolic flux analysis experiment. e, ECAR metabolic flux analysis of ITK-SYK-expressing T cells FACS sorted for eGFP+ from the spleens of acutely induced ITK-SYKCD4-creERT2 mice. As indicated in (d), the mice received anti-PD-L1 or isotype control antibodies on days one, three and five via intraperitoneal injection (n = 3 biological replicates per group). Data was normalized using total cellular protein. ECAR, extracellular acidification rate. P=two-sided Student’s t-test. The data are shown as box plots, in which the middle line denotes the median, the top and bottom box edges denote the 0.25 and 0.75 quantiles, respectively, and the whiskers denote the minimum and maximum values. f, OCR metabolic flux analysis from the same experiment as in (e) OCR, oxygen consumption rate. P=two-sided Student’s t-test. b, Representative data from a single mouse within the experiment shown in Fig. 2c, d. c, Representative results from 5 independent experiments with three biological replicates per group. e,f, Representative data from two independent experiments with three biological replicates per group.
