Figure 1

Stable DEAR1-KD in HMECs results in increased numbers of mammosphere independent of SMAD3 expression. (a) Representative bar plots showing mean ± SD counts of primary mammospheres (> 50um) from control vector (CshR) and DEAR1-KD (DshR) MCF10A cells across three trials. Each cell line was grown in at least 8 wells of 96-well plate in mammosphere growing conditions for 17 days for each individual trial. P-values were calculated using a two-tailed Student t-test between groups. **** p < 0.0001. (b) Representative images of mammospheres from (a) taken using a Zeiss microscope at ×10 magnification (Scale bars equal 100 µm). (c) Bar graphs representing the mean ± SD number of mammospheres (> 50 µm) formed from two control vector clones (CshR) and three DEAR1-KD clones (DshR) from 76N-E6 cells grown in mammosphere-growing conditions for 14 days across three trials. P-values were calculated using Wilcoxon Rank Sum tests. **p < 0.01 and ****p < 0.0001. (d) Representative images at ×10 magnification of CshR1 (left) and DshR3 (right) mammospheres quantified in panel (c) in the absence (top) and presence (bottom) of TGF-β (Scale bars equal 100 µm). (e) Bar plots showing the mean ± SD number of mammospheres formed from CshR, DshR, DshR/shC, and DshR/shSM3 cells without (left) and with (right) TGF-β treatment across 3 trials. p-values calculated using Wilcoxon Rank Sum tests. *p < 0.05–0.1 and ****p < 0.0001. (f) Bar graphs represent the mean ± SD number of mammospheres (> 50 µm) formed from two control vector clones (CshR) and three DEAR1-KD clones (DshR) from 76N-E6 cells treated with TGF-β and grown in mammosphere growing conditions for 14 days across three trials. P-values were calculated using Wilcoxon Rank Sum tests. *p < 0.05–0.1 and ****p < 0.0001. The barplots in (c), (e) and (f) were created using RStudio (Version 1.0.143), https://www.rstudio.com.