Figure 5
The potential role of GDF/BMP ligands in AP differentiation. (A) ATM Genes regulated by cold exposure. *P < 0.05, **P < 0.01, ***P < 0.001 by Student’s t-test. (B) The expression of BMP receptor and Smad genes in AP. (C) Western blot analysis in sorted CD45−/CD31− cells from HFD-fed VAT treated with the indicated ligand at 500 ng/ml for 30 min. HSP90 expression serves as loading control (N = 5). (D) qPCR analysis of Scd1 and Ppargc1a expression in FACS sorted AP from HFD-fed VAT after adipocyte differentiation. Cells were differentiated for 14 days in the presence or absence of the indicated ligand at 250 ng/ml. (E) GAL-ELK1 luciferase assay in HEK293T cells to monitor RET stimulation. Cells were transfected to express the appropriate luciferase reporter construct and an appropriate GFRA coreceptor as indicated with (right) or without (left) RET. Transfected cells were stimulated with the indicated ligand. Results shown as mean fold induction ± SEM (N = 3). (F) Expression of Gfra and Ret receptor genes in warm and cold conditions in FACS sorted ATM1, ATM2, and AP. Data are shown as mean ± SEM (N = 3).
