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. 2009 Sep 16;284(46):31726–31734. doi: 10.1074/jbc.M109.050914

FIGURE 5.

FIGURE 5.

Knockdown of Insig-2a expression by Insig-2a siRNA selectively promotes SREBP-1c processing. A, rat hepatoma cells were transfected with or without Insig-2a siRNA or control siRNA targeting GAPDH, and total RNA was isolated. The mRNA abundance of Insig-2a, Insig-2b, and Insig-1 was measured by real-time PCR analysis as described under “Materials and Methods”; 18 S rRNA was used as the control. The data are expressed as -fold change in the expression of Insig mRNA compared with mRNA levels in untransfected cells. B, rat hepatoma cells were co-transfected with pHisSREBP-1cFLAG plus either control siRNA for GAPDH or Insig-2a siRNA for 48 h, and cells were harvested. Nuclear extracts were separated by SDS-PAGE and immunoblotted for anti-His or anti-histone H1 antibodies. A representative blot of microsomal proteins fractionated by SDS-PAGE and immunoblotted using anti-Insig-2 antibodies is shown. The slowly migrating band above His-nSREBP-1c in the right two lanes represents the phosphorylated form of His-nSREBP-1c.