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. 2024 Feb 14;10(7):eadi1736. doi: 10.1126/sciadv.adi1736

Fig. 2. RAS-induced EMP and transformation capabilities are ZEB1-dependent.

Fig. 2.

(A and C) HME-RASER-miR200c cells (RASER-miR200c) or HME-RASER-empty cells (RASER-empty) after 28 days of induction with 4-OHT (IND). (D and F) HME-RASER-CRISPR ZEB1 clones (RASER-ZEB1#1, RASER-ZEB1#2, and RASER-ZEB1#3) or HME-RASER-CRISPR scramble (RASER-SCR) cells after 28 days of 4-OHT treatment (IND). (A) and (D) Flow cytometry analysis and quantification of CD24−/low/CD44+ cells. Data are presented as median ± range of four independent experiments (n = 4). (B and E) Transformation potential analysis by soft agar colony formation assay. The number of colonies (as defined by >20 cells) is indicated. Data are presented as median ± range of (B) four independent experiments in duplicate (n = 8), or (E) three independent experiments in duplicate (n = 6). (B) Images of GFP-positive colonies or (E) phase-contrast colonies (scale bars, 200 μm). (C) and (F) Quantification and phase-contrast images of mammospheres (scale bars, 200 μm). Data are presented as median ± range of four independent experiments (n = 4). P values are calculated by one-way ANOVA, Tukey multiple comparisons test (****P ≤ 0.0001).