Journal: Oncogene

Article Title: KMT2C mediates the estrogen dependence of breast cancer through regulation of ERα enhancer function

doi: 10.1038/s41388-018-0273-5

Figure Lengend Snippet: KMT2C loss suppresses ERα target gene expression. a Supervised analysis of the 7938 differentially expressed genes between shRenilla and shKMT2C (shKMT2C#1 and #2 combined) MCF7 cells. All cells are cultured in full serum containing media. b GSEA showing 3857 genes downregulated in shKMT2C cells are enriched among genes downregulated following 5-day estrogen deprivation of shRenilla cells (≥3-fold). c GSEA of 3857 genes downregulated in shKMT2C as compared to the Hallmark Estrogen Response Early Geneset (Broad Institute). d GSEA of 3857 genes downregulated in shKMT2C as compared the Hallmark Estrogen Response Late Geneset. e mRNA levels; values correspond to the mean of three replicates ± s.e.m. E2, estradiol. f mRNA levels; values correspond to the mean of three replicates ± s.e.m.; two-tailed Student’s t -test with a desired FDR = 1% was used to determine statistical significance; ** P < 0.01, *** P < 0.001, **** P < 0.0001. Data correspond to one representative assay from a total of two or three independent assays. g Immunoblot; β-actin used as loading control. h mRNA levels, as measured by qRT-PCR, from RNA taken from the mammary glands of 12-week old virgin females. Values correspond to the mean of three replicates ± s.e.m. i Representative slides of PR expression in mammary glands from 12-week-old virgin females

Article Snippet: All cells are cultured in full serum containing media. b GSEA showing 3857 genes downregulated in shKMT2C cells are enriched among genes downregulated following 5-day estrogen deprivation of shRenilla cells (≥3-fold). c GSEA of 3857 genes downregulated in shKMT2C as compared to the Hallmark Estrogen Response Early Geneset (Broad Institute). d GSEA of 3857 genes downregulated in shKMT2C as compared the Hallmark Estrogen Response Late Geneset. e mRNA levels; values correspond to the mean of three replicates ± s.e.m.

Techniques: Targeted Gene Expression, Cell Culture, Two Tailed Test, Western Blot, Control, Quantitative RT-PCR, Expressing

Journal: Oncogene

Article Title: KMT2C mediates the estrogen dependence of breast cancer through regulation of ERα enhancer function

doi: 10.1038/s41388-018-0273-5

Figure Lengend Snippet: KMT2C loss promotes hormone independent outgrowth. a Cells were assayed for proliferation in CSS media using the alamarBlue viability assay. Values correspond to the mean of six experimental replicates ± s.e.m. Data correspond to one representative assay from a total of two independent assays. b Supervised analysis of the 8622 differentially expressed genes between shKMT2C#2 cells in CSS media for 5 days and shKMT2C-R. CSS, charcoal stripped media. c Supervised analysis of the 9398 differentially expressed genes between shRenilla cells in CSS media for 5 days and shKMT2C-R. d GSEA of 4347 genes significantly downregulated in b as compared to the Hallmark Estrogen Response Early Geneset. e GSEA of 4559 genes significantly downregulated in c as compared to the Hallmark Estrogen Response Early Geneset. f MCF7 shKMT2C-R cells were treated with DMSO, estradiol (E2), fulvestrant (Fulv.), AZD9496 (9496), ARN1917 (1917), GDC927 (927), or RU58668 (58668) at indicated doses. Proliferation assayed using the alamarBlue viability assay. Values correspond to mean of 6 experimental replicates ± s.e.m. g Percent of cases with mutations detected in patient samples (MSKCC ) compared to those in TCGA Luminal A/B . h Progression free survival for ER+ breast cancer patients on single agent aromatase inhibitor (AI)

Article Snippet: All cells are cultured in full serum containing media. b GSEA showing 3857 genes downregulated in shKMT2C cells are enriched among genes downregulated following 5-day estrogen deprivation of shRenilla cells (≥3-fold). c GSEA of 3857 genes downregulated in shKMT2C as compared to the Hallmark Estrogen Response Early Geneset (Broad Institute). d GSEA of 3857 genes downregulated in shKMT2C as compared the Hallmark Estrogen Response Late Geneset. e mRNA levels; values correspond to the mean of three replicates ± s.e.m.

Techniques: Viability Assay