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. 2022 Oct 13;31(19-20):604–620. doi: 10.1089/scd.2021.0279

FIG. 6.

FIG. 6.

ASC adipogenesis is independent of breast cancer subtype, while fibril ECM production is dependent on breast cancer subtype. (A, B) ASCs were stimulated with MCF-7 and MDA-MB-231CM for 3 days followed by culture in adipogenic medium for 7 days. Oil Red O staining revealed no difference in adipogenesis between cohorts. N = 3 biological replicates and N = 2 technical replicates (C) qRT-PCR analysis of MCF-7 and MDA-MB-231 stimulated ASCs revealed significantly elevated expression (*P < 0.05) of PPAR-γ in MCF-7 CM stimulated ASCs. MCF-7 and MDA-MB-231 stimulated ASCs also exhibited significantly lower expression of UCP-1 compared to the control. N = 3 biological replicates and N = 2 technical replicates, error bars = SEM (D) ASCs were initially stimulated with MCF-7 CM and estrogen, maintained by alternating CM and full culture medium for 2 weeks, and stained with Sirius Red/Fast Green for fibril collagen and noncollagenous protein content. Significantly increased (*P < 0.05) fibril collagen content was observed in MCF-7 CM stimulated ASCs in week 1 but was not observed in TNBC CM stimulated ASCs at either timepoint. (E) Further treatment of MCF-7 cells and ASCs with 17β-estradiol revealed significantly elevated (*P < 0.05; **P < 0.01) expression of fibril collagen in MCF-7 cells only. N = 3 biological and technical replicates, error bars = SEM. LEP, Leptin; PPARγ, peroxisome proliferator activated receptor gamma; UCP1, uncoupling protein 1.