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. Author manuscript; available in PMC: 2021 May 11.
Published in final edited form as: Nature. 2020 Nov 11;588(7839):693–698. doi: 10.1038/s41586-020-2911-7

Extended Data Fig. 6. Additional data on the characterization of lymphocyte infiltration into murine tumors.

Extended Data Fig. 6.

a. Immunofluorescence staining (left panel) and quantitative estimate (right panel) of CD45+ leukocytes in control and PCSK9KO tumors grown in syngeneic C57BL/6 mice. Scale bar = 50 μm. n=3 biologically independent samples. Four fluorescent fields for each of the three samples were counted. Error bars, mean ± S.E.M. P value calculated using unpaired two-sided t-test. b.Immunofluorescence staining (left panel) and quantitative estimate (right panel) of CD8a+ cells in control and PCSK9KO B16F10 tumors. Scale bar = 20 μm. n=3 biologically independent samples. Four fluorescent fields for each of the three samples were counted. Error bars, mean ± S.E.M. P value calculated using unpaired two-sided t test. c. Quantitative estimates of CD4+ and 452 CD8+ T cells in the spleens of mice bearing control and PCSK9KO B16F10 tumors as determined by flow cytometry. n=3 mice per group. Error bars, mean ± SEM. P values calculated using unpaired two-sided t-test. d. Flow cytometry determination of the percentage of intratumoral CD8+ T cells that were IFNγ+. n=6,5 tumors in the two groups. Error bars, mean ± S.E.M. P value calculated by unpaired two-sided t-test. e, f. Q-RT-PCR analysis of intratumoural IFNG (e) and GZMB (f) mRNA levels in control and PCSK9KO tumors. n=3 and 4 tumors for INFG and GZMB groups, respectively. Error bars, mean ± S.E.M. P values were determined by unpaired two-sided t test. g-i. Flow cytometry characterization of the cell surface expression levels of exhaustion markers for intratumoural CD8+ T cells in vector control and PCSK9KO tumors. n=6, 5 tumors. Error bars, mean ± S.E.M. P values were determined by unpaired two-sided t test. j. Evolocumab and anti-PD1 treatment schedule for syngeneic 4T1 tumor model. k. Growth of 4T1 tumors treated with anti-PD1 and/or evolocumab. n=5 mice per group. P values were determined by two-way ANOVA test. l. Kaplan-Meier survival curve for mice in k. P values were determined by log-rank test. m. Frequency of CD8+ T cells in 4T1 tumors treated with anti-PD1 and/or evolocumab. n=5 tumors per group. Error bars, mean ± SEM. P values were determined by unpaired two-sided t test. n. Frequency of IFNγ+CD8+ T cells in 4T1 tumors treated with anti-PD1 and/or evolocumab. n=5 tumors per group. Error bars, mean 469 ± SEM. P values were determined by unpaired two-sided t test.