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. 2009 Dec;11(12):1318–1328. doi: 10.1593/neo.91084

Figure 1.

Figure 1

Cell surface expression of CD44 and CD24 in Twist-dysregulated breast cells. (A) Immunoblots showing the downregulation of Twist in MCF-10A/Twist, MCF-7/Twist, and MDA-MB-231 breast cancer cell lines by short hairpin RNA against Twist delivered using lentiviral vectors. Lower panels show Twist transient expression levels in MCF-10A and MCF-7 cell lines expressing Twist by retroviral delivery. The antibody against Twist was generated in-house and was validated. Actin was scored as loading control. (B) Flow cytometry analysis of various cell lines for CD44 and CD24 expression. Unstained and single-antibody stained cells were used as controls for setting the quadrants. The events in the lower right quadrant represent the CD44+/CD24-/low subpopulation. The analyzed data are represented as dot plots of (first row) immortalized normal mammary epithelial cell line MCF-10A and breast cancer cell lines MCF-7 and MDA-MB-231, (second row) stable Twistoverexpressing cell lines MCF-10A/Twist and MCF-7/Twist, (third row) transiently transduced Twist-expressing MCF-10A and MCF-7, and (fourth row) Twist knockdown in MCF-10A/Twist, MCF-7/Twist, and MDA-MB-231 breast cancer cells. Ten thousand viable cells were gated for each dot plot acquisition. Results are representative of five separate experiments.