Extended Data Fig. 6: MEN1 and MLL1 regulation of cytokine-related genes expression and immune cell infiltration, related to Fig. 3.
(a) Western blot showing control (sgCtrl), cGAS knockout (left; sgcGAS-1, sgcGAS-2) and MAVS knockout (right; sgMAVS-1, sgMAVS-2) A549 cells. Vinculin was used as a loading control. (b) Western blot of TBK1 and p-TBK1 in control (sgCtrl) and MEN1 knockout (sgMEN1-1, sgMEN1-2) A549 cells. Vinculin was used as a loading control. (c) Xenograft tumor growth curve in immunodeficient mice inoculated with control (sgCtrl), cGAS and MAVS knockout (sgcGAS, sgMAVS) A549 cells. Each data point represents mean ± s.e.m. tumor volumes (n=10 in sgCtrl, sgMEN1-1, and sgMEN1-2 group). Two-way ANOVA test was used for the statistical test of the growth curves. (d) Heatmap of upregulated genes (p < 0.01 and Log2(FC) > 1) in the leukocyte migration term in MEN1 knockout A549 xenograft tumors. (e) GO analysis of differentially expressed mouse genes in MEN1 knockout versus control A549 tumors. Neutrophil related terms are highlighted. Wald tests defined in DEseq2 were used to calculate the p values. (f) Representative IHC images showing the infiltration of neutrophils for control (sgCtrl) and MEN1 knockout (sgMEN1-1, sgMEN1-2) A549 xenograft tumors. (g) Bar plot showing enriched KEGG terms of 478 upregulated genes from menin inhibitor MI-389 treated MV4-11 human MLL leukemia cells. X-axis represents the number of genes. Wald tests defined in DEseq2 were used to calculate the p values. (h) Dot plot showing enriched GO terms of 478 upregulated genes from menin inhibitor MI-389 treated MV4-11 human MLL leukemia cells. Wald tests defined in DEseq2 were used to calculate the p values. (i) Percentage of infiltrated Neutrophil cells in A549 xenograft tumors with and without knockout of MEN1 or in combination with MAVS and/or cGAS knockout. Mean ± s.e.m of quantifications from 10 tumor IHC sections biological replicates were shown (unpaired two-tailed Student's t-test). *: p value <0.05.
