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. Author manuscript; available in PMC: 2017 Feb 19.

Published in final edited form as: Biochem Biophys Res Commun. 2016 Jan 20;470(4):783–791. doi: 10.1016/j.bbrc.2016.01.064

Fig. 1 — (a) Western blot analysis was performed to determine protein expression of TRB3 in the tissues from 12-week-old male C57BL/6. (b) Brown preadipocytes were differentiated to brown adipose cells for 8 days. Cells were collected at indicated days to determine TRB3 mRNA expression using Real-time PCR analysis (n=5). (c) Brown preadipocytes were infected with retrovirus containing empty vector (pBabe) or TRB3. Western blot analysis was performed to detect TRB3 expression in the cells. (d–j) Brown preadipocytes had undergone differentiation for 8 days. On day 8, the cells were fixed and stained with Oil Red O to visualize fat accumulation (d and e; Bar 100 µm). RNA was isolated from cells during differentiation to measure mRNA expression of PRDM16 (f), CIEDA (g), PPARγ (h), and UCP1 (i) (n=5). Protein lysates were subjected to gel electrophoresis to determine the UCP1 expression (j). Data are the means ± S.E.M. * indicates p<0.05, ** indicates p<0.01, and *** indicates p<0.001 vs. basal in the same group. # indicates p<0.05, ## indicates p<0.01, and ### indicates p<0.001 vs. corresponding control.

Fig. 1 — (a) Western blot analysis was performed to determine protein expression of TRB3 in the tissues from 12-week-old male C57BL/6. (b) Brown preadipocytes were differentiated to brown adipose cells for 8 days. Cells were collected at indicated days to determine TRB3 mRNA expression using Real-time PCR analysis (n=5). (c) Brown preadipocytes were infected with retrovirus containing empty vector (pBabe) or TRB3. Western blot analysis was performed to detect TRB3 expression in the cells. (d–j) Brown preadipocytes had undergone differentiation for 8 days. On day 8, the cells were fixed and stained with Oil Red O to visualize fat accumulation (d and e; Bar 100 µm). RNA was isolated from cells during differentiation to measure mRNA expression of PRDM16 (f), CIEDA (g), PPARγ (h), and UCP1 (i) (n=5). Protein lysates were subjected to gel electrophoresis to determine the UCP1 expression (j). Data are the means ± S.E.M. * indicates p<0.05, ** indicates p<0.01, and *** indicates p<0.001 vs. basal in the same group. # indicates p<0.05, ## indicates p<0.01, and ### indicates p<0.001 vs. corresponding control.