Fig 2: SRC-3 aggregates are associated with microtubules, redistribute PML and SUMO and compromise nuclear membrane integrity. (A) 3D deconvoluted model constructed as described in Materials and Methods using Z-stacks acquired with confocal microscopy from SRC-3-transfected cells and immunostained for SRC-3 (red) and β-tubulin (green) and imaged. Blue is DAPI (100× magnification). (B) Immunofluorescence of β-tubulin (green) and SRC-3 (red) showing regions of increased perinuclear tubulin density in SRC-3 overexpressing cells. Nuclear protrusion of SRC-3 and DNA (blue) are indicated with arrows (63× magnification). (C) SRC-3 expression interferes with mitotic microtubule dynamics. MCF-7 cells were infected with Ad-LacZ or Ad-SRC-3 on coverslips and 48 h later were treated with taxol for 18hrs and IF performed to detect mitotic microtubules and SRC-3 (63× magnification). (D) Cells treated in C were enumerated for the presence of mitotic figures. Bars are S.D., ** p < 0.01. (E) Z-stack images of cells transfected with SRC-3-YFP and immunostained for lamin B1 (red) and tubulin (green) immunofluorescence showing nuclear membrane distortion, and degradation associated with cytoplasmic SRC-3 aggregates (100× magnification). (F) IF of lamin B1 in SRC-3-GFP-transfected cells showing cells with discontinuous nuclear membranes, lamin B1 fragments, and distortions of nuclear shape.