Fig. 2.

The expression of Irx3 was correlated with the beige/brown adipocyte associated genes in vitro.

(A–G) In fully differentiated beige/brown adipocytes from preadipocytes isolated from IWAT, EWAT, and BAT of C57BL/6J and 129/Sv mice, Ucp1 mRNA expression was correlated with Pgc-1α (A) and Dio2 (B). Irx3 mRNA expression showed positive correlation with mRNA levels of (C) Ucp1, (D) Pgc-1α, (E) Dio2, and (F) Cox7a1. Irx3 mRNA expression showed no correlation with Leptin (G). Cells from C57BL/6J were shown in red, and from 129/Sv were in blue. Cells from IWAT, EWAT, and BAT were shown as dot, triangle, and square, respectively. The correlation was analyzed using ΔCT. (H-J) (H) Irx3 (I) Ucp1 and (J) Pparγ mRNA expression during the time course of beige adipocyte differentiation cultures (n = 3). Quantitative amounts of gene expression were normalized to the housekeeping gene 36b4. (K–L) Irx3 and Ucp1 protein levels at different time points during beige adipocyte differentiation from IWAT SVFs (K), during brown adipocyte differentiation from BAT SVFs (L). (M) Protein expression of Irx3 and Ap2 at different time points during white adipocyte differentiation from IWAT SVFs. (N) Intracellular location of Ucp1 and Irx3 protein in the fully differentiated beige adipocytes. Ucp1 protein, Irx3 protein and nucleus were indicated in green, red, and blue, respectively. Scar bar, 20 μm. Data were presented as mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001. The results are representative of at least three independent experiments.