Fig. 7. The mAKAP complex contributes to NFATc1 activation. (A) Primary myocytes were co-transfected with an expression vector for Flag-tagged NFATc1 and either the control (a,b) or mAKAP (c,d) siRNA expression plasmid. The myocytes were treated for 2 days with no drug, 10 µmol/l Iso, 100 µmol/l PE, 400 nmol/l CsA and/or 100 nmol/l KT5720. Representative cells cultured in Iso-containing medium are shown. Cells were fixed and stained with antibodies for the Flag-tag (green, a,c) and -actinin (red, b,d) and with Hoechst 33258 DNA stain (blue, b,d). Bar, 20 µm. (B) The fraction of transfected myocytes with predominately nuclear NFATc1 (mean±s.e.m.). ^*P<0.05, ^**P<0.002 in comparison to the similarly treated control siRNA sample, n6; ANOVA, P<10^-4 for entire data set and Iso- and PE-treated data alone.