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. 2021 Jun 15;148(12):dev199281. doi: 10.1242/dev.199281

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© 2021. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 4. — Laminin α5 is essential for pregnancy-induced growth and differentiation of the mammary gland. (A) Schematic showing the experimental outline of tamoxifen induction of K8-CreERT2 before pregnancy. Representative images of carmine alum-stained #4 mammary glands of 17.5 dpc pregnant Lama^fl/fl;- and Lama^fl/fl;K8-CreERT2 animals. (B) Representative H&E images of 17.5 dpc pregnant Lama5^+/+;K8-CreERT2 and Lama5^fl/fl;K8-CreERT2 mice. (C) Representative immunofluorescence images of 17.5 dpc pregnant Lama5^+/+;K8-CreERT2 and Lama5^fl/fl;K8-CreERT2 glands immunostained with Pan-laminin and E-cadherin, or K8 and K14 antibodies. Square shows area of inset. (D) Representative immunofluorescence images of 17.5 dpc pregnant Lama5^+/+;K8-CreERT2 and Lama5^fl/fl;K8-CreERT2 glands immunostained with ZO-1 or MUC1, and E-cadherin antibodies. (E) Quantification of the alveolar diameter of 17.5 dpc (+/+, n=3; fl/fl, n=3 individuals analyzed) and PP2 mice (+/+, n=4; fl/fl, n=3 individuals analyzed). N=3-4 individual animals. Data points represent mean alveolar diameter quantified from individual animals. (F) Representative immunofluorescence images of PP2 mammary glands immunostained with K14 or Pan-laminin and E-cadherin antibodies. (G) Representative immunofluorescence images of PP2 mammary glands immunostained with ZO-1 or MUC1, and E-cadherin antibodies. Data are mean±s.d. **P<0.01 (unpaired two-tailed Student's t-test). Scale bars: 10 mm (A); 200 µm (B); 50 µm (C,D,F,G).