Fig. 6

Changes in stem cell-related phenotypes of breast cancer cells following altered expression levels of SIX1. A Representative images of mammospheres and their corresponding mammosphere-formation efficiencies in 66cl4-SCR, 66cl4-shSix1 KD1, 66cl4-shSix1 KD2, MCF-7-NC, and MCF-7-SIX1 cells. The images were captured at a magnification of × 100, and the scale bar represents 200 µm. The data are presented as the mean ± standard deviation (SD) (n = 3). B Flow cytometry analysis of CD24 and CD49f expression in 66cl4-SCR, 66cl4-shSix1 KD1, and 66cl4-shSix1 KD2 cells, as well as CD24 and CD44 expression in MCF-7-NC and MCF-7-SIX1 cells. The red boxes indicate the stem cell population. C Bar graph showing the activity of ALDH enzyme in 66cl4-SCR, 66cl4-shSix1 KD1, 66cl4-shSix1 KD2, MCF-7-NC, and MCF-7-SIX1 cells. Higher OD values indicates stronger activity (n = 3). D Cell growth of 66cl4-SCR, 66cl4-shSix1 KD1, 66cl4-shSix1 KD2, MCF-7-NC, and MCF-7-SIX1 cells cultured for 0, 24, 48, 72, and 96 h (n = 4). Statistical significance is denoted as *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 respectively, compared to the control group