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. Author manuscript; available in PMC: 2011 Dec 16.
Published in final edited form as: Nature. 2010 Dec 16;468(7326):933–939. doi: 10.1038/nature09564

Figure 4.

Figure 4

CRTC3 attenuates cAMP signaling in adipose. A. cAMP content in WAT (top) and cultured MEFs (bottom) from wild-type and CRTC3−/− mice. Exposure to FSK indicated. (**; P<0.01) B. Effect of CRTC3 over-expression (top) or RNAi mediated knockdown (bottom) on cAMP accumulation in wild-type MEFs exposed to FSK as indicated. (***; P<0.001) C. Top, Q-PCR analysis showing relative mRNA amounts for RGS2 in primary cultured adipocytes (WT, CRTC3−/−) exposed to FSK as indicated. Bottom, Q-PCR analysis of RGS2 mRNA amounts in WAT from NC or HFD-fed mice (WT, CRTC3−/−). (*; P<0.05. **; P<0.01) D. Effect of RGS2 over-expression (top) or RNAi mediated knockdown (bottom) on cAMP accumulation in wild-type MEFs exposed to FSK as shown. (*; P<0.05. ***; P<0.001) E. Top, transient assay of RGS2-luc reporter activity in HEK293T cells exposed to FSK. Effect of CRTC3 over-expression or RNAi-mediated depletion indicated. Bottom, chromatin immunoprecipitation (ChIP) assay showing occupancy of CRTC3 over the RGS2 promoter in MEFs exposed to FSK as indicated. F. Top, immunoblot showing relative amounts of wild-type and S72N variant epitope-tagged CRTC3 polypeptides in cytoplasmic and nuclear fractions from transfected HEK293T cells. Fractionation of control cytoplasmic (ACC) and nuclear (CREB) proteins shown. Bottom, transient transfection assay of HEK293T cells showing relative activities of wild-type and S72N CRTC3 expression vectors co-transfected with RGS2-luc reporter plasmid. Exposure to FSK indicated.