Figure 5 from A RAS(ON) Multi-Selective Inhibitor Combination Therapy Triggers Long-term Tumor Control through Senescence-Associated Tumor-Immune Equilibrium in Pancreatic Ductal Adenocarcinoma
CD4 T cells prevent outgrowth of residual disease. A, Scheme of experimental design (KPC1 orthotopic transplant into wild-type C57Bl/6 mice). B, Percentage change in tumor volume compared with day 0, as measured by weekly ultrasound. The y-axis indicates the percentage change in tumor volume, and the x-axis indicates the time point after treatment initiation. Each dot represents an individual mouse. Statistical testing: Two-way ANOVA with multiple comparisons (within each time point, comparing treatment groups with every other treatment group) and correcting for multiple comparisons with a Tukey test. All statistically significant comparisons are shown. C, Representative hematoxylin and eosin (H&E) images of residual tissue following 42 days of indicated treatment. D, Scheme of experimental design for E–H (KPC1-ZsGreen orthotopic transplant into wild-type C57Bl/6 mice). E, Fraction of CD8+ T cells out of total CD45 cells by flow cytometry at 7 days after the initiation of indicated treatments. See F for the legend. Statistical testing for E–G: Ordinary one-way ANOVA with multiple comparisons, comparing the means of each treatment group with every other treatment group and correcting for multiple comparisons using a Sidak test. Only relevant statistically significant comparisons are shown. F, Fraction of CD4+ T cells out of total CD45 cells by flow cytometry at 7 days after the initiation of indicated treatments. Each dot represents an individual mouse. G, Fraction of tumor cells (%) positive for MHC-II by flow cytometry at 7 days after the initiation of indicated treatments. See F for the legend. Each dot represents an individual mouse. Statistical testing: Ordinary one-way ANOVA, comparing the means of every treatment group with every other treatment group and correcting for multiple comparisons with a Sidak test. All statistically significant comparisons are shown. H, Fraction of CD4 T cells (%) positive for indicated cytotoxicity and activation of Th1 markers by flow cytometry 7 days after treatment initiation. Statistical testing: Two-way ANOVA with multiple comparisons, comparing the mean of each treatment group with every other treatment group within the relevant time point and correcting for multiple comparisons with a Tukey test. Only statistically significant comparisons are shown. I, Representative IF images of residual tissue following 42 days of indicated treatments (see scheme in A) staining for pan-CK, Ki67, cleaved caspase 3, and DAPI. (A and D, Created with BioRender.com.)