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. 2019 Aug 2;38(18):e101426. doi: 10.15252/embj.2018101426

Figure 2. C/EBPα inhibits hormone‐induced expression of stem cell markers and acts as a cell growth modulator in vivo .

Figure 2

  1. T47DindC/EBPα cells induced or not with Dox were untreated or exposed to R5020 for 24 h, and the expression of CD44 and CD24 was measured using specific antibodies (CD44‐APC and CD24‐P, respectively) and fluorescence‐activated cell sorting analysis. Data are represented as mean and SD from three experiments performed in duplicate. The P‐values were obtained by ANOVA followed by Tukey test.
  2. Left panel: Scheme of the Mouse Intra‐Ductal (MIND) xenograft approach: tumor cells expressing luciferase are injected intraductally via the teat (Sflomos et al, 2016). Seven days after injection, CEBPα expression is induced with 2 mg/ml doxycycline in the drinking water maintained throughout the experiment. Tumor growth is assessed by bioluminescence (IVIS). Right panel: Tumor growth of T47DindC/EBPα‐MIND marked with luciferase (Sflomos et al, 2016) treated or not with Dox was assessed by bioluminescence. Results are shown as means ± SEM. Five mice were used for each condition. In total, 17 and 19 glands were analyzed from control and C/EBPα. P‐values were obtained by Student's t‐test and were calculated relative to control animals (*P ≤ 0.05)
  3. Upper panels: T47DindC/EBPα and T47DyindC/EBPα (PR‐) cells expressing inducible recombinant C/EBPα were exposed to Doxycycline (1 μg/ml) for different times, and then, the number of cells was monitored. Data are represented as mean ± SD from two experiments performed in triplicate. P‐values were obtained by Student's t‐test and were calculated relative to untreated (−DOX). The numbers in parenthesis correspond to the average of the ratio +DOX over −DOX for each time point. Lower panels: T47D‐ and T47Dy‐inducible C/EBPα cells were treated with doxycycline (1 μg/ml Dox) for 24 h, and the expression of C/EBPα was monitored by immune fluorescence microscopy (left) and Western blot (right). The Western blot confirms that T4Dy cells do not express PR. HP1γ was used as loading control. The band with a molecular weight around 55 kDa correspond to the TetO‐C/EBPα ds Tomato fusion protein.

Source data are available online for this figure.