Fig 1: MiR-200c restoration alters the profile of cytokines secreted by aggressive mammary and breast carcinoma cells to favor those that enhance antitumor macrophage polarization.a A cytokine array was conducted on Met-1 TripZ-200c cell conditioned medium (CM; mouse) and a V-PLEX cytokine assay was conducted on BT549 TripZ-200c cell CM (human). Depicted are cytokines of interest that were altered in both models after miR-200c restoration and are known to promote monocyte infiltration, activation, or differentiation. b Met-1 (top) and BT549 (bottom) cells were harvested after 96 and 48 h, respectively, transient miR-200c restoration. qRT-PCR was conducted for CSF2 (GM-CSF), CXCL10, and CCL2, mean mRNA expression relative to GAPDH for 3 experiments conducted in triplicate ± s.d., N = 8–9, Student’s unpaired two-tailed t test. c qRT-PCR was conducted on Met-1 TripZ-200c tumors from Fig. 2d that were treated ± Dox for 24 to 30 days. Shown is the mean mRNA expression relative to Gapdh ± s.e.m, N = 6–8, Students unpaired two-tailed t test. d GM-CSF expression was observed in the CM of Met-1 cells after miR-200c restoration for 96 h via transient transfection. Shown is the mean cytokine expression (pg/mL) for three experiments completed in triplicate ± s.d., N = 9, Students unpaired two-tailed t test. e RAW264.7 macrophages were treated with 50 ng/mL or 65 ng/mL GM-CSF for 48 h. Cells were harvested for mRNA extraction via TRIzol and macrophage polarization was determined via qRT-PCR for an M1 gene Nos2 (nitric oxide synthase/iNOS) and a cytokine secreted by M1 macrophages Tnfa (tumor necrosis factor-a, TNFa). Representative mean mRNA expression relative to Gapdh for one experiment of three ± s.d., N = 3, Student’s unpaired two-tailed t test.