Extended Data Fig. 7: Characterization of senescent cells in culture and in vivo.
From: The efficacy of chemotherapy is limited by intratumoral senescent cells expressing PD-L2
(a) p21 staining in pellets from Panc02 cells, WT and PD-L2-KO, untreated or treated with doxorubicin (200 nM, 48 h, evaluated at day 7, n = 4 experiments). Scale bar = 100 μm. (b) GSEA plots for representative gene sets associated to cellular senescence in doxorubicin-treated versus untreated WT Panc02 cells, as well as in doxorubicin-treated (as described) versus untreated PD-L2-KO Panc02 cells at day 7 after treatment. DESEq2 was used for differential expression analysis with fold change shrinkage as implemented in the lfcShrink function. Functional enrichment analysis was performed over gene sets defined in the Molecular Signatures Database (MSigDB) hallmark gene set collection. Data from 4 biological replicates. (c) Absolute quantifications of intratumoral levels of CCL2 and interleukin 6 measured by a commercial multiplexed system with beads bound to antibodies, in WT and PD-L2-KO tumors, untreated or treated with doxorubicin (4 mg/kg, days 7, 10 and 24) and anti-Gr1 (200 µg per injection, as described for Fig. 3e), or the same dose of IgG isotype control. N = 5 mice per experimental condition. 1 way ANOVAs with Tukey post-tests were applied. (d) Relative levels of intratumoral cytokines and chemokines in WT and PD-L2-KO tumors, untreated or treated with doxorubicin (4 mg/kg, on days 7, 10 and 24, as described). N = 5 mice per experimental condition.
