Cells with stable expression of Hugl1 shRNA (MH11), Hugl2 shRNA (MH2C) and control shRNA (MC) were cultured in Matrigel for 21 days, fixed with 2% PFA, permeabilized, and incubated with primary antibodies marking apical (anti-GM130, red), cytoskeletal (anti-phalloidin, yellow), or basal (anti-Laminin V, green) domains followed by fluorescently labeled secondary antibodies (FITC, Alexa 488, or Alexa 647). Slides were mounted with antifade mounting media containing DAPI. Acini were imaged with a Leica confocal microscope to obtain equatorial cross sectional images of their morphology. (A–I) All images were taken at 630X using internal zoom. Subsections of B and E (white boxes) magnified in C and F. Scale bars indicate size in microns.