Fig. 2
From: A miR-327–FGF10–FGFR2-mediated autocrine signaling mechanism controls white fat browning
Identification of miR-327 and FGF10 as SVF-derived modulators of early browning. a Heatmap of significantly (P < 0.05) up- and downregulated miRNAs from 3-day CL-316243-treated visWAT-SVFs compared to vehicle-treated controls (n = 5 mice per sample; n = 2 samples per group). b Heatmap of significantly downregulated growth factor (GF)-targeting miRNAs extracted from a. c–f qPCR analysis of miR-327 in c, d 3-day CL-316243-treated SVFs or e, f 1-week 4 °C-treated SVFs of scWAT, visWAT, and iBAT compared to respective controls. Sno-202 served as an internal control (n = 5 samples per group). g Alignment details of miR-327 to Fgf10 3′ UTR binding site 1 (bs1) and binding site 2 (bs2). Xs mark nucleotides in bs1 and bs2 that were mutated to generate Mut-bs1 and Mut-bs2. h Quantification of Renilla luciferase activity normalized to Firefly luciferase activity in miR-327 mimic-treated HEK293T cells co-transfected with psiCHECK-2 vector, psiCHECK-2-Fgf10 bs1, psiCHECKTM-2-Fgf10 bs2, psiCHECKTM-2-Fgf10 Mut-bs1, or psiCHECKTM-2-Fgf10 Mut-bs2 (n = 5 samples per group). i Volcano plot of the FGF ligand and receptor families from genome-wide microarray analysis of 3-day CL-316243-treated visWAT-SVF. Lowest P-value is highlighted in red and highest fold-change is highlighted in blue (n = 4 mice per sample; n = 3 samples per group). j–m qPCR analysis of Fgf10 in j, k 3-day CL-316243-treated SVF or l, m 1-week 4 °C-treated SVF of scWAT, visWAT, and iBAT compared to respective controls (n = 5 samples per group). NS, not significant. *P < 0.05, **P < 0.01, and ***P < 0.001 by Student’s t-test. Data presented as mean ± s.e.m.
