VAPB expression enhances spheroid size and proliferation in mammary epithelial cells.

<p>(A) VAPB was stably expressed in MCF10A-HER2 cells via retroviral transduction, as judged by western blot analysis. (B) MCF10A-HER2-VAPB spheroids were cultured in three-dimensional Matrigel. Spheroid size was quantified and presented as average pixel area per spheroid (p<0.01, ANOVA). (C) Cell proliferation in spheroid culture was determined by immunofluorescence using an anti-PCNA antibody (red). Cell nuclei were stained blue by TO-PRO-3 nuclear stain. PCNA-positive nuclei were quantified from an average of 15 random spheroids for each of two experiments (p<0.05, t test). Arrows: PCNA positive cells.