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. 2021 Jul 15;9:691050. doi: 10.3389/fcell.2021.691050

FIGURE 3.

FIGURE 3

BMPR1a deficiency resulted in precocious alveolar differentiation during pregnancy. (A) Immunohistochemistry staining for WAP in control and cKO mammary glands at pregnancy day 14.5. n = 3 mice per group. Scale bar, 50 μm. (B) Immunohistochemistry staining and western blotting for β-Casein in control and cKO mammary glands at pregnancy day 14.5. n = 3 mice per group. β-Tubulin was used as a loading control. Scale bar, 50 μm. Statistical analysis the expression of β-Casein/β-Tubulin. n = 3 mice. (C) qRT-PCR analysis of Wap, Lalba and Csn2 in mammary epithelial cells isolated from control and cKO mice at pregnancy day 14.5. n = 4–6 biological replicates. (D) Immunofluorescence staining and western blotting for Plin2 in control and cKO mammary glands at pregnancy day 14.5. n = 3 mice per group. GAPDH was used as a loading control. Scale bar, 25 μm. Statistical analysis the expression of Plin2/GAPDH. n = 3 mice. (E) Heatmap for milk protein-related gene expression in control and cKO mammary glands at pregnancy day 14.5. n = 4 mice per group. (F) Immunohistochemistry staining and western blotting for Stat5 and pStat5 in control and cKO mammary glands at pregnancy day 14.5. n = 3 mice per group. β-Actin was used as a loading control. Scale bar, 50 μm. Statistical analysis the expression of pStat5/β-Actin. n = 3 mice. (G) Immunohistochemistry staining for Prlr in control and cKO mammary glands at pregnancy day 14.5. n = 3 mice per group. Scale bar, 50 μm. (H) qRT-PCR analysis of Elf5, Tnfsf11, Src, Socs-1, and Socs-2 in mammary epithelial cells isolated from control and cKO mice at pregnancy day 14.5. n ≥ 3 biological replicates. Data were presented as means ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.