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Fig. 4. Senescent mouse fibroblasts stimulate mammary epithelial branching morphogenesis. Primary organoids were embedded in collagen and plated in the upper chambers of transwells. Presenescent (Presen) or senescent (Sen) mBF were plated in the transwell lower chambers. Error bars show s.e.m. of triplicate wells. Shown is one of four experiments, all of which gave similar results. (A) Morphology and color coded explanation of branching classification of organoids cultured with presenescent (i and iii) or senescent (ii and iv) mBF. Shown are the core areas (white), and the primary (1ary, blue), secondary (2ary, yellow) and tertiary (3ary, red) branches. (B) Average organoid size (Total), length of branches (Branches) and core areas (Core) of organoids co-cultured with presenescent or senescent mBF. (C) Average number of primary (1ary), secondary (2ary) and tertiary (3ary) branches of organoids co-cultured with presenescent or senescent mBF. (D) Average length of primary (1ary), secondary (2ary) and tertiary (3ary) branches of organoids co-cultured with presenescent or senescent mBF. (E) Average number of epithelial nuclei, quantified by DAPI fluorescence, in organoids co-cultured with presenescent or senescent mBF.





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