Fig. 6: ER+ BC cells with the acetylation-defective 6KR mutant MED1 exhibit enhanced tumorigenesis and tolerance to hypoxic and oxidative stresses.
From: MED1 IDR deacetylation controls stress responsive genes through RNA Pol II recruitment
a, Elevated growth rate of 6KR/KO versus WT/KO MCF7 cells monitored by CellTiter Blue assay (n = 4 biological replicates). Data points report the mean ± s.e.m. The P value from a one-way analysis of variance (ANOVA) comparing two curves is shown. b, More colonies formed by 6KR/KO versus WT/KO MCF7 cells stained with crystal violet following 10 days of growth after initial seeding. c–e, Declined viability of 6KR/KO and WT/KO MCF7 cells treated with various concentrations of DFOM (24 h) (c), H2O2 (24 h) (d) or paraquat (48 h) (e) monitored by MTT assays showing tolerance of 6KR/KO (n = 4 biological replicates in each group). Bars report the mean ± s.e.m. If <0.05, the P values of a two-sided unpaired t-test comparing cell types are shown. f,g, Enhanced growth of tumors formed from 6KR/KO versus WT/KO MCF7 cells implanted into inguinal mammary fat pad in NSG mice. Images of the mice with orthotopic tumors (18 and 16 tumors for WT/KO and 6KR/KO cells, respectively) (f) and curves of tumor volume (g) are shown. Data points report the mean ± s.e.m. The P value from a one-way ANOVA comparing two groups before 95 days after implantation is indicated. h, Poly(A) RNA-seq for 6KR/KO and WT/KO MCF7 tumors developed in NSG mice. Three tumor samples (biological replicates) were examined for each type of tumor. A volcano plot is presented of the adjusted P value (two-sided) and FC for each detected gene, with the cutoff for defining DEGs (FC > 2, adjusted P < 0.05) indicated. i–k, GSEA showing that the upregulated genes in hypoxia-exposed (data from GSE153557) (i), paraquat-treated (data from GSE241672) (j) or H2O2-treated (data from study described above) (k) versus control MCF7 cells were enriched in the set of upregulated genes of 6KR/KO versus WT/KO MCF7 tumors. Adjusted P values are noted in addition to the NES. l,m, Increased lung metastasis of 6KR/KO versus WT/KO MCF7 cells in NSG mice with implantation of these cells originally into inguinal mammary fat pad. l, Representative hematoxylin and eosin staining images of lung sections within which metastatic foci are indicated. Scale bar, 100 μm. m, Quantification of metastatic foci. Data are the mean ± s.e.m. and the P value of a Wilcoxon rank-sum test (two-sided) is shown.
