Fig. 7: Overexpression of MEG8 decreases the stemness properties of the cells in breast cell lines.

A Measurement of MEG8 expression levels by RT-qPCR in the mammospheres and the total extract of T47D, MDA-MB-231 and MDA-MB-468 cell lines. B Percentage of holoclones, meroclones and paraclones generated by control and MEG8 overexpressing breast cell lines seeded at low density during 15 days. C Number of mammospheres formed comparing control and MEG8 overexpressing breast cell lines. D Percentage of mammospheres formed from one single cell of control and MEG8 overexpressing breast cell lines separated by FACS. E Quantification of the percentages of CD44 + CD24- cells in control and MEG8 overexpressing breast cell lines by FACS. F Quantification of the percentage of NANOG + CD24- cells in control and MEG8 overexpressing breast cell lines by FACS. G Measurement of BMI1, SOX9 and KLF4 expression levels by RT-qPCR in control and MEG8 overexpressing breast cell lines. H Expression levels of Sox9 and cMyc genes by a microarray analysis in the breast tissue of three different strains of mice, C57BL/6 J, SAMP6 and AKRJ. We used 3/4-, 16- or 93-weeks (w) old animals. Here, we only show the results of females (n = 3 in each condition). I Tumorigenicity of tumorspheres in vivo. Mammospheres from T47D and MDA-MB-468 control and MEG8 cells were injected in nude mice (N = 8). Mice inoculated with the T47D cell line were treated with 4 mg/mL of β-estradiol. Graphs represent the tumor size (mean ± SEM). The figures show the average of three independent experiments performed in triplicate. Data were analyzed using Student’s t-test. *p < 0.05; **p < 0.01; ***p < 0.001.