Extended Data Fig. 5: Ouabain eliminates liver preneoplastic senescent cells.
a, Representative images of immunofluorescence staining of Nras. Mice were treated with vehicle (n = 9) or ouabain (n = 12) as explained in Fig 4a. Nras is stained in red. Scale bar, 70 μm. b, Immunofluorescence staining and quantification of p21Cip1 in Nras-positive senescent hepatocytes vs Nras-negative normal hepatocytes. Nras is stained in green, p21Cip1 is stained in red. White arrows indicate Nras-positive, p21Cip1-positive cells; green arrow indicates a Nras-positive, p21Cip1-negative cells (n = 10 per group). c, SCID/beige mice were treated with saline or Digoxin (1mg/kg) on two consecutive days, 5 days after hydrodynamic transduction of Nras-GFP. Mice were culled 6 hours after the second treatment. Representative images of immunofluorescence staining of GFP and cleaved caspase-3 and quantification of intensity levels in 1/2 independent experiments (n = 200 cells). Scale bar, 50 μm. Statistical significance was calculated using unpaired two-tailed Student’s t-test. Data represent mean ± s.d; n represents number of mice.
