Loss of Elf5 leads to increased EMT gene expression programs
(a) Heat map representation of microarray data displaying the expression of several key EMT-related genes in wild type and Elf5-KO mammary glands on lactation day 1. (b) GSEA data showing the enrichment of four published EMT gene signatures38-41 in Elf5-KO mammary glands as compared to wild type glands. NES: normalized enrichment score. (c, e) Western blot analyses of HA-Elf5 and E-cadherin protein levels in NMuMG cells transduced with vector (control) or HA-Elf5. Uncropped images of blots are shown in Supplementary Fig. S9. (d) Phase contrast and E-cadherin immunofluorescence images of NMuMG (control) or NMuMG-Elf5 cells undergoing TGFβ-induced EMT when cultured in low cell density. Experiments using cells cultured in high density are shown in Supplementary Fig. S1i. Size bar = 100 μm for brightfield and 40 μm for E-cadherin. (f) qRT-PCR analysis of expression of EMT-related genes in NMuMG (control) or NMuMG-Elf5 cells, with or without TGFβ treatment. Real time PCR values were normalized to the housekeeping gene Gapdh. Experiments were performed three times, each with qRT-PCR in technical duplicate, and data presented as the mean ± SD. * p < 0.05 by Student’s t-test.