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. 2015 Mar 3;34(11):1493–1508. doi: 10.15252/embj.201489004

Figure 1.

Figure 1

Shp2 ablation or inhibition inhibits self-renewal and induces senescence in cultured PyMT tumor cells
  1. Scheme of Shp2 ablation in cultures of PyMT;Shp2fl/fl tumor cells using retroviruses, which expressed CreERT2-GFP or control GFP.
  2. Western blot analysis of Shp2 in control and Shp2 ko cells.
  3. Immunofluorescence analysis of Ki67 on control and Shp2 ko cells at day 7. Scale bar, 50 μm.
  4. Quantification of the numbers of Ki67+ cells shown in (C). Error bars represent SEM (n = 3). **P < 0.01.
  5. Sphere formation by control and Shp2 ko cells at day 7. Scale bar, 200 μm.
  6. Quantification of the numbers of spheres formed by control and Shp2 ko cells shown in (E). Error bars represent SEM (n = 3). **P < 0.01.
  7. Growth kinetics of spheres formed by control and Shp2 ko cells by counting sphere cells at day 4, 7, and 12. Error bars represent SEM (n = 3).
  8. SA-β-gal staining on spheres formed by control and Shp2 ko cells at day 7. Scale bar, 100 μm.
  9. Quantification of the numbers of senescent cells in the spheres shown in (H). Error bars represent SEM (n = 3). **P < 0.01.
  10. Western blot analysis of H3K9me3, p27, p53 pS18 (Ser18-phosphorylated p53), total p53, Shp2, and α-tubulin in control and Shp2 ko cells.
  11. SA-β-gal staining on spheres formed by control cells and those treated by the Shp2 inhibitor GS493 at 15 μM at day 7. Scale bar, 100 μm.
  12. Quantification of the numbers of senescent cells shown in (K). Error bars represent SEM (n = 3). **P < 0.01.
  13. Quantification of the numbers of spheres formed by control cells and those treated by the Shp2 inhibitor GS493 at 15 μM. Error bars represent SEM (n = 3). **P < 0.01. Statistical significance was assessed by Student's unpaired t-test.