Figure 2: MYC causes Reversible Transcriptional Repression of Antitumor Immune Responses.

A. Heat map shows similarity between MYC-driven murine HCC (MYC-On, n=3 vs. MYC-Off, n=3) and 11 human HCC transcriptome data sets including the TCGA cohort; the top 10 upregulated and downregulated genes with enrichment z-scores across the different data sets are shown. Bar plot at the bottom shows log of p-value of similarity between the individual cohorts and murine MYC-HCC.

B. Heatmap shows z-scores of enrichment of canonical pathways and biological functions in murine MYC-HCC transcriptome and the human HCC immune signatures of exhausted PD1+ CD8 T cells(31) and inactivated 41BB-ve PD1+ T cells in human HCC(30).

C. Gene signature derived from murine MYC HCC used to classify human HCC in the TCGA cohort to identify MYC-driven human HCCs (created using Biorender.com).

D. Human HCCs enriched in mMYC^On signature show MYC molecular pathway activation and strong association with MYC gene amplification.

E. Human HCCs with high expression of mMYC^On signature showed enrichment of a gene program of T cell exclusion and resistance to anti-PD1 checkpoint therapy (32), and upregulation of CTLA4.

F. PCA plot comparing the transcriptional profile of a transgenic model of HCC overexpressing wild-type MYC (MYC^On, n=16), a mutant MYC defective in Miz1 binding (V394D) (MYC^Vd, n=11) and tumors upon MYC inactivation for a short time point of 16 hours (MYC^Off, n=8)(36) to identify MYC/Miz-1 repressed genes.

G. Heatmap shows genes which are reversibly repressed by MYC in the MHC protein complex, complement system and cytokines.

H. Immunohistochemistry shows the abundance of macrophages (F4/80+) by day 1 and CD4 T-cell recruitment by day 4 of MYC inactivation in HCC.

I. Immune deconvolution analysis showing comparison of abundance of different immune subsets in murine MYC HCC upon MYC inactivation.

**p<0.01, ***p<0.0001.