Figure 7. PKA is required for the formation of long-range, inter-genic interactions between the PPARγ2 promoter and (A) the adiponectin promoter, (B) the leptin promoter, (C) the perilipin 1 promoter, (D) the perilipin 2/ADRP promoter.

The EF1α promoter (E) is shown as a control. 3T3-L1 cells infected with retroviral vectors expressing control shRNA or shRNA targeting the PKA-Cα or PKA-Cβ subunits were generated (see Methods). 3C assays were performed on these cells at 0 or at 6 h post-differentiation in the presence of complete differentiation cocktail. Interaction frequencies were quantified as described in the legends for Figures 1–2. Results indicate the average of three independent experiments +/− standard deviation. Values for the 6 h samples treated with control shRNA were compared to values for the 6 h samples treated with PKA-Cα or PKA-Cβ shRNA by one-tailed t-test. **p<0.01. (F) Oil Red O staining of infected 3T3-L1 cells allowed to differentiate for 4 days. (G) Western blots indicating the levels of the PKA-Cα or PKA-Cβ subunits in the infected cells. PI-3K levels were monitored as a loading control.