Figure 5.

HCG-induced increases in the binding of SREBP to the LRBP promoter was abrogated on pretreatment with miR-122 antagomir. LNA-conjugated miR-122 antagomir was injected into the bursa of the ovaries of superovulated rats on day 4, followed by hCG on day 5. Ovaries were collected 4 hours later and were processed for ChIP assay as described in detail in Materials and Methods. Upper panel, Tissues were cross-linked, and ChIP assay was performed with mouse IgG (negative CTL) or Pol II antibody (positive CTL), as described in Materials and Methods. The purified DNA samples from ChIP assay (positive and negative CTLs) as well as input DNA were amplified by PCR using rat primer sequences for the GAPDH gene promoter, and the PCR products were analyzed using 1.2% agarose gel electrophoresis. Lower panel, The immunoprecipitated DNA, along with input DNA, was subjected to real-time PCR quantitation using specific primers and probes for MVK promoter and GAPDH. The graph represents MVK promoter levels in the immunoprecipitated samples normalized to input DNA and shown as fold change vs CTL. Error bars represent mean ± SE. *, P < .05 vs CTL and **, P < .05 vs hCG; n = 3.