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Fig. 3. mAKAP is involved in the induction of adrenergic-stimulated myocyte hypertrophy. (A) Following co-transfection with a GFP expression vector and an expression vector for either control (left panels) or mAKAP siRNA (right panels), primary myocytes were cultured for 2 days in minimal medium containing either no drug (a-f), 10 µmol/l Iso (g-l), or 100 µmol/l PE (m-r). Cells were stained with mAKAP VO56 (red, b,e,h,k,n,q) and -actinin (blue, c,f,i,l,o,r) antibodies. The nuclei of transfected, GFP-expressing myocytes (green) are indicated with arrowheads on the mAKAP panels. Bar, 20 µm. (B) The level of mAKAP expression in myocytes infected with control or mAKAP siRNA-expressing adenoviruses were assayed by immunoblotting with VO56 mAKAP antibody. Total protein was detected by Ponceau S stain; the major band is myosin. (n=3) (C) Mean cell surface area (± s.e.m.) for GFP-labeled cells co-expressing either the control siRNA, the mAKAP siRNA, or the mAKAP siRNA and exogenous, myc-tagged mAKAP protein (mAKAP rescue). ^*P=0.04, ^**P=0.01, ^***P=0.003 for the samples compared, n>150 with myocytes derived from 7 independent cultures; ANOVA, P=0.004 and 0.02 for Iso- and PE-treated data, respectively.

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