Fig. 2

Differentiation potential of LGSCs in vitro. a, b Heatmap of RNA-sequence analysis of LGSCs cultured for 5, 7, 10, and 14 days. a Heatmap of cell division genes; b heatmap of cell differentiation and gland development; n = 3. c qRT-PCR analysis of adult stem cell and differentiated markers of LGSCs cultured for 5, 7, 10, and 14 days. Secretory cell marker AQP5, secretory protein gene of LG Ltf, and ductal cell marker Krt19 are significantly upregulated. Adult stem cell marker Krt14 is significantly downregulated, ***, P < 0.01; n = 3. d Immunofluorescent staining of LGCSs cultured for 5, 7, 10, and 14 days. As elongating culture time, cells expressing Krt14 (red) and Ki67 (red) are significantly decreased, while cells expressing Krt19 (red) emerge, scale bar, 50 μm. Nuclear staining, DAPI (blue). e, f Flow cytometer analysis of Krt14 expression of LGSCs cultured for 5, 6, 7, 8, 9, 10, and 14 days. e Flow cytometric profile, M1 (Krt14⁻), M2 (Krt14^Low), M3 (Krt14^High); f comparative graph of LGSC subpopulations shows that LGSCs cultured for 7 days have the highest proportion of Krt14-positive cells