Extended Data Fig. 2: Analysis of the tumor immune infiltrate in transgenic Pten-/-; Trp53-/- orthotopic model.

a) Bar graphs showing results of the FACS analysis on the Pten-/-; Trp53-/- orthotopic model. Same strategy as shown in Extended Data Fig. 1. Events are gated on CD45+ cells. b-c) % of naive, central memory, effector, effector memory and Treg cells gated on the (b) CD4+ subset and the (c) CD8+ subset. d-e) % of cells expressing functional markers gated on CD4+ and CD8+ cells. f) % of CD39+ cells gated on CD8+ cells. g) Heat map illustrating top 200 differentially expressed genes from bulk mRNA-Seq on sorted macrophages from the Pten-/-; Trp53-/- orthotopic model. h) Experimental scheme of in vitro conditioning of macrophages. i) Representative FACS plot showing non- conditioned macrophages (Untreated=Untr, left) and macrophages exposed to conditioned media from Pten-/-; Trp53-/- cells (Conditioned =CM-tr, right). j) Proliferation of CD8 + T cells exposed to supernatant derived from ex vivo macrophages. Macrophages were sorted from either tumor (n = 3) or healthy tissues (n = 6) and cultured for two days to collect conditioned supernatant. CD8 + T cells were subsequently exposed to these supernatants to assess their proliferation: bar graph represents the number of divisions. k) % of ARG1+ cells on CD206 + FACS-sorted macrophages (Untr n = 7, CM-tr n = 12. L) RT-qPCR gene expression analysis on CD206+ and CD206- FACS sorted macrophages (n = 3). Statistical analyses were performed using two-tailed unpaired Student’s t test. Values are presented as mean ± SEM. Schematic in h created using BioRender.com.