Fig. 5. Involution is severely affected in the mutant mammary glands. (A) HE staining of mammary glands sections. (Left panels) Mammary glands from control mice. (Right panels) Mammary glands from mutant mice. (Upper panels) Day 1 of involution; (second panels) day 3 of involution; scale bar: 40 µm; (third panels) Carmine Red-stained whole mounts of mammary glands at day 21 of involution; scale bar: 800 µm; (bottom panels) HE staining of sections from the corresponding whole-mounts samples; scale bar: 80 µm. (B) TUNEL analysis of control (upper left panels) and mutant (upper right panel) involuting mammary glands 3 days post parturition. TUNEL-positive cells are in green. Lower panels show DAPI staining of the corresponding sections. Scale bar: 40 µm. (C) Quantification of TUNEL-positive cells (left). Ten 200x magnification fields of view were randomly counted. The apoptosis index was calculated as following: ([number of TUNEL positive cells]/[total number of cells])x100. The total cell numbers per 400x magnification fields of view was also quantified and graphed (right panel). Statistical analysis was performed using Student’s t-test. Error bars represent standard errors of the mean.

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