Fig. 5: FOXM1 assembles an epigenetic repressor complex to inhibit STING and consequently unfolded protein response pathways to control stress ligand ULBP1 expression in cancer cells. | Nature Communications

Fig. 5: FOXM1 assembles an epigenetic repressor complex to inhibit STING and consequently unfolded protein response pathways to control stress ligand ULBP1 expression in cancer cells.

From: Epigenetic silencing of DNA sensing pathway by FOXM1 blocks stress ligand-dependent antitumor immunity and immune memory

Fig. 5

a Heatmap showing the log-normalized expression of top genes from GSEA pathway enrichment analysis. Genes representing pathways with the highest enrichment in E0771 Foxm1 knockout tumors compared to scrambled control are shown. b Western blots of cGas-STING pathway proteins in scrambled (Ctrl) and FOXM1 knockout E0771 (left) and MDA-MB-231 cells (right). β-actin or GAPDH was used as a loading control. The arrow indicates the band for total STING. c, d qRT-PCR analysis of STING (c), and DNMT1 and UHRF1 mRNA (d) in MDA-MB-231 Ctrl and FOXM1 KO cells. e Western blot analysis of DNMT1 and UHRF1 in MDA-MB-231 and E0771 Ctrl and FOXM1 KO cells. f ChIP-qPCR in MDA-MB-231 cells showing FOXM1 enrichment on the DNMT1 promoter. g Western blot analysis of MDA-MB-231 and BT549 cells treated with DNMT1 inhibitor (GSK-3484862) using antibody against STING. h Western blot analysis of MDA-MB-231 transfected with scrambled (siCtrl) or siDNMT1 cells using antibodies against cGas-STING pathway proteins. i ChIP-qPCR in MDA-MB-231 cells showing DNMT1 enrichment on STING promoter. j ChIP-qPCR in MDA-MB-231 Ctrl and FOXM1 KO cells showing enrichment of H3K4me3 on STING promoter. k MethyLight qRT-PCR assay using primers spanning STING promoter in Ctrl and FOXM1 KO MDA-MB-231 cells. The bar graph shows the relative log2 of percentage STING promoter methylation in FOXM1 wild-type and FOXM1 knockout groups. l Western blot analyses in scrambled and Foxm1 KO E0771 cells using antibodies against the indicated UPR proteins. m qRT-PCR on Foxm1 KO cells transfected with scrambled or Sting siRNA using primers against MHC-I, IFNβ, and ULBP1. All bar graphs represent log2 of fold change in gene expression or fold enrichment on promoters. All qPCR data represent the mean +/− SEM of three biological replicates with two technical replicates per group. Western blots are representative of three independent experiments. p-values for panels (c, f, i–k) were calculated using a two-sided Welch’s t test. p-values for (d and m) were calculated using multiple t test with Benjamini, Krieger, and Yekutieli correction. Control and FOXM1 knockout groups in all bar graphs are represented by dark blue circles or red squares, respectively. Source data are provided as a Source Data file.

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