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. 2014 Feb 4;5:3224. doi: 10.1038/ncomms4224

Figure 7. NRDc coexists with PGC-1α at the UCP1 enhancer.

Figure 7

(a) Western blot analysis of cytosolic and nuclear fraction of mature brown adipocytes treated for 2 h with or without isoproterenol (ISO) by indicated antibodies. Full blots are presented in Supplementary Fig. 9. (b) Mature brown adipocytes stained with anti-NRDc antibody and 4',6-diamidino-2-phenylindole were visualized by confocal microscopy. Scale bars, 20 μm. (c) ChIP with anti-PGC-1α antibody followed by qPCR demonstrates that Nrd1 knockdown in mature brown adipocytes increases PGC-1α recruitment to PPRE of the UCP1 enhancer. Results were normalized with input DNA, and are shown as the ratio to that in the control miR-treated cells (IP: αPGC-1α). n=5. (d) ChIP with anti-NRDc antibody followed by qPCR demonstrates that ISO treatment decreases NRDc binding to PPRE of the UCP1 enhancer. Results are shown as the fold induction relative to that in cells without ISO treatment (IP: Control IgG). n=4. (e) ChIP/re-ChIP analysis was performed with anti-NRDc and PGC-1α antibodies followed by qPCR targeting the PPRE of the UCP1 enhancer. n=3. All data represent means±s.e.m. *P<0.05, (two-tailed Student’s t-test).