Fig. 4

BMP4 is required for normal acinus formation in Matrigel 3-D culture. a, b Matrigel 3-D culture of 344SQ-NTC and Bmp4-KD cells (#2 and #3). Phase contrast (a) and confocal microscope (b) images of 3-D acini were taken 12 days after cell seeding. Blue: DAPI, red: β-catenin, green: ZO-1. c Matrigel 3-D culture of 344SQ_vec and miR-200 cells. Confocal images of 3-D acini were taken 12 days after cell seeding. Blue: DAPI, red: β-catenin, green: ZO-1. d Matrigel 3-D culture of 344SQ-NTC and Bmp4-KD cells (#2 and #3). Confocal images of 3-D acini were taken 7 days after cell seeding, and spindle angles of dividing cells were measured using ImageJ (http://imagej.nih.gov/ij/). Blue: DAPI, red: α-tubulin. White arrow: direction of cell division; yellow arrow with asterisk: spindle angle. Box-and-whisker plot denotes median and upper/lower quartiles + 1.5 × IQR. p, two-tailed Student’s t-test