Increased nuclear localization of maspin^D346E correlates with increased HDAC1 interaction.

Abstract

<p>(<b>A</b>) Confocal imaging of endogenous maspin (green) and HDAC1 (red) immunofluorescence staining in MCF10A cells. Scale bar = 20 µm (<b>B</b>). Bimolecular fluorescence complementation (BIFC) of live PC3 and DU145 cells transiently transected with pBiFC maspin^WT YC or pBiFC maspin^D346E YC in combination with pBiFC HDAC1 YN. The BiFC of bJunYN and BiFC bFosYC and BiFC of bJunYN and BiFC bFosYC Δ179–193 were used as positive and negative controls, respectfully, in PC3 cells. Scale bar = 10 µm (<b>C</b>) Immunofluorescence imaging (x40) of live PC3 cells after co-transfection with either pEGFP N1-maspin^WT or pEGFP N1 maspin^D346E in combination with pcDNA3.1 HDAC1-RFP. → indicates cytosolic maspin/HDAC1 interaction and ▸ indicates nuclear maspin/HDAC1 interaction. Scale bar = 20 µm. (<b>D</b>) Western blot of recombinant maspin and HDAC1 after immunoprecipitation (IP) with maspin antibody in DU145 cells. The mouse IgG was used as a negative control. Total levels of maspin, HDAC1, and loading control GAPDH are shown in the input panel. Numbers below represent normalized HDAC1/maspin ratio.</p