Fig. 4: Mammary organoids exhibit robust regeneration capability.

a Colony number of cultured basal cells in serial passages. Ctrl: control basal cell, wnt3A: Wnt3A treated basal cell, M2: basal cell co-cultured with M2. n = 3 replicates, *P < 0.05, **P < 0.01, ***P < 0.001, unpaired t-test. b The proportion of colonies and organoids formed in basal cells co-cultured with M2 in serial passages. n = 3 replicates. c The representative organoid images formed after 4 passages, stained for K14 and K8. Scale bar: 100 μm. d Organoids show a strong mammary reconstitution rate in transplantation experiments compared with that of colonies and the representative images of the reconstituted mammary glands on the right of the panel. Scale bars: 5 mm, 1 mm. e Volcano plot of the differentially expressed genes in organoids produced by co-culture with M2 compared with that of control (FC > 2 and adjust P value < 0.05). f Venn diagram of up-regulated overlapping genes in DEGs (differentially expressed genes) in M2 co-culture and wnt3A treated basal cells. g GO analysis of overlapping genes in up-regulated DEGs in M2 co-cultured and wnt3A treated basal cells. h GO analysis of up-regulated DEGs in M2 co-cultured basal cells. i GO analysis of up-regulated DEGs in wnt3A treated basal cells. j GSEA shows that compared with the control cells, M2 co-cultured cells were enriched with gene signatures of MaSC. k GSEA shows that compared with the control cells, M2 co-cultured cells were enriched with gene signatures of mammary regeneration.