Fig. 7

Chronic TE defects confer resistance to anti-tumor immune attack and immunotherapy in vitro and in vivo. a Heatmap of relative expression (left), and immunoblots for proteins (right), of some key inflammatory response pathway genes in control and flavopiridol-treated cells. b Control and flavo pre-treated cells were stimulated with IFN-α, IFNγ or TNF-α for 30 mins or FasL (c) for 24 h, and the readout was measured with the indicated proteins by immunoblotting (b) or viability assay (c). d Relative viability of B16/F10-OVA cells co-cultured with activated CD8 + CTLs (1:1 ratio) isolated from the spleens of OT-I mice. P: Welch two-sample t-test, e in vivo assessment of resistance to NK-mediated anti-tumor response: mice are injected intravenously with 2 × 10⁵ B16/F10-OVA cells pre-stained with CFSE, and tumor cell seeding in the lungs is assessed 1 h later by measuring OVA mRNA levels in the lungs by qPCR (middle) or by flow cytometry for CFSE + cells (right). ΔNK: NK cell depletion by subcutaneous pre-injection of mice with anti-asialo GM antibody 1 day prior to tumor cell injection. The p-value is for Welch two sample t-test. f Tumor growth of control or flavopiridol pre-treated CT26 mouse carcinoma cells in syngeneic (Balb/c) mice treated with control (IgG) or anti-CTLA4 antibody (9H10), P: Welch two-sample t-test. g Weights and images (bottom) of excised tumors of indicated treatment groups after 21 days post-injection. h Immunoblots of some of the excised tumors from the treatment groups in experiment in f for H2-K^D (HLA-I), β2m, and Actin. Error bars: standard deviations of 3 (c), 2 (d), 6 (e), and 4 (g) replicates