Fig 1: Manf stimulated browning via p38 MAPK signaling. (A) qPCR analysis of thermogenic genes in primary iWAT incubated with CM from primary hepatocytes infected with Ad-GFP or Ad-Manf for 48 h (n = 3). (B) Thermogenic gene expression in primary iWAT treated with recombinant Manf and Fgf21 (n = 3). (C) Phosphorylated of p38 and ATF2 in primary adipocyte from iWAT treated with different concentration of Manf for 30 min. (D) Phosphorylated and total p38 and ATF2 in primary adipocyte from iWAT incubated with 2.5 nM Manf for different periods of time. (E) Primary adipocytes were pretreated with SB203580 (10 µM) for 2 h, then Manf (2.5 nM) for an additional 48 h. mRNA levels of thermogenic genes were detected by qPCR (n = 3). (F) Phosphorylated and total p38 and ATF2 protein levels in iWAT from WT and Tg mice fed an HFD for 12 wk (n = 3). Each experiment was independently performed two to three times. All data are mean ± SEM. *, P < 0.05; **, P < 0.01. Ctrl, control.