Figure 6. Nuclear AKT interacts with cofilin.

(A) A2058 cells were serum starved for 24 h and then stimulated (+) or not (−) with 10% serum for 30 min, as indicated. Nuclear fractions were immunoprecipitated (IP) with anti-cofilin or control IgG antibodies. Cell lysates prior to immunoprecipitation were used as input. Input and co-IP proteins were resolved on SDS-PAGE and immunoblotted for the detection of p-AKT⁴⁷³, total AKT, β-actin and cofilin. (B and C) A2058 cells were double immunostained for: (B) AKT (green) and Cofilin (red); (C) cofilin (green) and β-actin (red). DAPI-stained nuclei are shown in blue. Immunostained cells were analyzed by confocal laser-scanning microscopy. Images are projections of one stack from the middle plane of the nucleus. Reconstructed orthogonal projections are presented as viewed in the xz planes, showing co-localized immunostaining in the nucleus (yellow pixels, arrowhead); scale bar, 5 μm.