Fig. 2: Clodronate liposomes lead to attenuation of mammary stem cell activity.

a Schematic illustration of the serial passages in 3D culture, and the mammary regeneration assay of basal cells. b CL treatment inhibits basal cell proliferation: graph shows numbers of basal cell colonies in each passage. n = 3 replications, *P < 0.05, **P < 0.01, n.s represents no significance, unpaired t-test. c CL treatment promotes luminal cell proliferation: graph shows numbers of luminal cell colonies in each passage. n = 3 replications, *P < 0.05, n.s represents no significance, unpaired t-test. d Mammary gland reconstitution efficiency of basal cells in transplantation. The mammary outgrowth numbers and sizes (shown as the percentage of fat pad filled) are combined from three independent experiments. **P < 0.01, unpaired t-test. Scale bar: 5 mm. e Volcano plot of the differentially expressed genes in basal cells from control and CL treated mice. (FC > 1 and adjust P value < 0.05). f GO (Gene Ontology) analysis of the basal cells for differentially upregulated gene in CL treated mice compared with that of control mice. g GSEA shows that compared with the control mice, the enrichment score of the gene signature of mammary regeneration in the basal cells in CL treated mice was lower. h GSEA shows that compared with the control mice, the enrichment score of the gene signature of MaSC (Procr labeled) in the basal cells in CL treated mice was lower. i GSEA shows that compared with the control mice, the enrichment score of the gene signature of MaSC (Bcl11b labeled) in the basal cells in CL treated mice was higher. j qPCR analysis shows that compared with that in the control mice, the expression of Bcl11b in basal cells of CL treated mice was increased, n = 3 replications, *P < 0.05, unpaired t-test. k t-SNE plot showing the basal cells of control mice and colored by relative expression (gray = low, purple = high) of Bcl11b. l t-SNE plot showing the basal cells of CL treated mice and colored by relative expression (gray = low, purple = high) of Bcl11b. (Ctrl: control mice, CL: CL treated mice).