Fig. 6. Genetic elimination of senescent CAFs in transplanted PDACs leads to increased rates of activated CD8+ T cells.

a Diagram of experimental design. 6422c1 KPC cells, which generate tumors with high endogenous p16+ CAF content, were injected into the pancreata of Ink-ATTAC mice. Mice were treated with AP20187 three times a week during tumor growth. b Sections of tumors formed by KPC cells in mice either untreated (UNT) or treated with AP, co-stained for p16 and with CK8 (top), marking epithelial cells or with Pdpn (bottom), marking CAFs. c Relative stromal p16+ area in tumors formed by the KPC cells, with or without AP treatment. Values were measured by image analysis and normalized to mean of controls. n = 4 tumors, >1 mm²/tumor. d Percentage of p16+ cells out of Pdpn+ CAFs in same samples, scored by image analysis. n = 5,4 tumors. e Tumor weights upon sacrifice, in mice treated with AP or untreated. n = 5,4 tumors. f Quantification of indicated immune cell populations in tumors formed in control and AP-treated mice, scored by FACS. n = 5,4 tumors. g Percentages of activated CD8+ T cells in same mice, scored with indicated markers by FACS. h Quantification of additional indicated immune cell populations in tumors formed in control and AP-treated mice, scored by FACS. n = 5,4 tumors. CD11b and CD11c mark monocytic populations, gMDSC – granulocytic myeloid-derived suppressor cells (Ly6G⁺Ly6C^low), mMDSC – monocytic myeloid-derived suppressor cells (Ly6G^-Ly6C^high), F4/80 marks macrophages, FoxP3 marks regulatory T cells. i Relative stromal p16+ area in tumors formed by FC1199 KPC cells, with or without AP treatment, measured by image analysis. n = 6,5 tumors, >1 mm²/tumor. j Tumor weights upon sacrifice, in mice transplanted with FC1199 KPC cells treated with AP or untreated. n = 6,5 tumors. k, l Quantification of indicated T cell populations in FC1199 tumors formed in control and AP-treated mice, scored by FACS. n = 6,5 tumors. All values indicate mean ± SEM. t test. Scale bars = 20 μm.