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. 2020 Mar 19;8:68. doi: 10.3389/fcell.2020.00068

FIGURE 1.

FIGURE 1

Lactation induction in primary mammary organoids. (A,B) FGF2 pretreatment increases lactation capacity of primary mammary organoids. (A) Scheme depicting the experimental design. BOM, basal organoid medium; LM, lactation medium; FGF2, FGF2 medium. (B) Expression of milk genes Csn2 and Wap in organoids treated with BOM, LM, or FGF2 followed by LM. The values are relative to BOM. The plot shows mean + SD; n = 2. One-way ANOVA, *p < 0.05. (C) Bright-field images and maximum intensity projection images from confocal imaging of whole-mount organoids after treatment with FGF2 only or with FGF2 followed by LM. Yellow-to-brown staining shows F-actin. Scale bars represent 100 μm. (D) Bright-field image and maximum intensity projection images from confocal imaging of whole-mount organoid treated with FGF2 followed by LM. Red, oil red O (lipids); green, F-actin; blue, DAPI (nuclei). Scale bars represent 100 μm. (E,F) Quantification of β-casein expression in organoids treated with FGF2, or FGF2 followed by LM. (E) RT-qPCR analysis of β-casein gene Csn2. The values are relative to FGF2. The plot shows mean ± SD; n = 3. Unpaired Student’s t-test, two tailed, ****p < 0.0001. (F) Western blot analysis of β-casein expression on protein level. The plot shows quantification of band density. The values are relative to FGF2. (G) Immunohistochemical staining of β-casein in organoids treated with FGF2 or FGF2 and then LM at days 6 and 10, respectively. Marked area is shown in higher magnification. Scale bars represent 100 μm.