Extended Data Fig. 10: Loss of Ccl2 and Ccl7 mimics loss of Qpctl in LL/2 tumors.
a,b, CRISPR/Cas9-edited Ccl2–/–Ccl7–/– LL/2 cells were generated. a, Quantification of MCPs in the cell supernatants (n = 1). b, WT mice were inoculated with WT or Ccl2–/–Ccl7–/– LL/2 cells. CCL7 quantification in plasma of naive and tumor-bearing mice (n = 5 (naive and day 14) and 10 (day 19)). *P = 0.008, **P < 0.0001. c,d, WT mice were inoculated with WT, Qpctl–/– or Ccl2–/–Ccl7–/– LL/2 cells. c, Tumor growth measurements (n = 10). Gray lines represent individual mice and overlay of fitting spline cubic curves for each group is shown. *P < 0.0001. d, Tumor weight at day 14 (n = 5). *P = 0.03, **P = 0.008. e,f,g, WT mice were inoculated as described in (c). e, Identification of Ly6C+ monocytic cells (red circle) and F4/80hi macrophages (blue circle) among tumor leukocytes (gated on live, singlets, CD45+CD11b+Ly6G–). f, Quantification of Ly6C+ monocytic cells and macrophages (n = 5). g, Frequency of F4/80+MHCII+ macrophages (n = 5). *P = 0.02, **P = 0.008. h, Qpctl+/+ and Qpctl–/– EO771 bearing mice were treated with Ctrl Isotype or anti-CD8 depletion antibody. Identification of CD8+ T cells among CD3+ T cells in blood, 2 days after antibody injection. i, Qpctl+/+ and Qpctl–/– littermates were inoculated with Qpctl+/+ or Qpctl–/– EO771 tumor cells, respectively, and treated with Ctrl Isotype or anti-PD-L1 blocking antibody. Frequency of tumor-associated GzmB+CD8+ T cells (n = 12 (Qpctl+/+) or 14 (Qpctl–/–)). NS, P > 0.05. Bars are medians and symbols represent individual mice. Data shown is representative (a,b,c,d,f,g,h) or pooled from 2 experiments (i). All experiments were repeated independently (≥ twice). Statistical analysis was done using unpaired, nonparametric Mann-Whitney U-test (b,d,g,i) or Two-way ANOVA with Tukey correction (c).
