Fig. 2.

Expression of G protein mRNAs in J774A.1 and silencing of G proteins by subtype-specific siRNAs. (A) RT-PCR was performed by using total RNA isolated from J774A.1. After PCR (30 cycles) with subtype-specific primers, one-fifth of the product was loaded on a 1% agarose gel. For controls (C), cDNA corresponding to the Gβ or Gα subunit was used. Alternatively, the RT product derived from the J774A.1 cells (J) was used. SM, size markers. (B and C) siRNA constructs contained in pSK vectors were transfected into HEK-293 cells with plasmid, maintaining the cDNA corresponding to the enhanced GFP fusion form of each G protein. After 48 h, Western blotting was performed with an anti-GFP antibody.