Fig. 1. Human adipocytes induce transcription of EMT genes in a co-cultured ER⁺ human breast cancer cell model.

(A and B) MCF7 cells were co-cultured for 5 days with insulin-sensitive (IS) adipocytes that were differentiated from human primary pre-adipocytes, or ex vivo-induced insulin-resistant (IR) adipocytes from the same source, and compared to control MCF7 alone (con). Expression of selected EMT genes was analyzed by commercial PCR array (A) and Ingenuity pathway analysis (IPA; B). Data are from 3 independent experiments. (C) Fluorimetric glucose uptake assay in primary adipocytes after 24 hours treatment with recombinant human 250 pM TNF-α,10 nM insulin, both, or neither to assess insulin sensitivity status. Data are mean ± SEM from 3 independent experiments. (D) Expression of SNAI1 and SNAI2 in MCF7 cells cultured with exosomes purified from insulin-sensitive (IS) or insulin-resistant (IR) adipocyte–conditioned media or with exosomes purified from conditioned media of matched, undifferentiated pre-adipocyte control cells (pre) compared to MCF7 cells alone (con). Fold-change in expression of selected mRNAs encoding Snail (SNAI1) and Slug (SNAI2) was measured by RT-PCR relative to RNA encoding β-actin (ACTB). Data are mean ± SEM from 3 independent experiments; * P <0.05, *** P <0.005, and ns not significant by unpaired two-tailed t-test (C and D).