Extended Data Fig. 6: Ouabain eliminates preneoplastic senescent cells.
a, Representative images of immunofluorescence staining of β-catenin (green) and synaptophysin (red) in tumoral pituitaries from 18.5 dpc Hesx1Cre/+;Ctnnb1lox(ex3)/+ mice that were cultured in the presence of either ABT-737 (2.5 μM), ouabain (250 nM and 500 nM) or vehicle (DMSO) (n = 10 per group). Scale bar, 50 μm. b, Quantitative analysis of the immunofluorescence in (a) demonstrates that ABT-737 and ouabain significantly reduce the number of β-catenin-positive cells. Statistical significance was calculated using Kruskal-Wallis and Dunn’s multiple comparisons test. c, Representative images of immunofluorescence staining of ACTH (adrenocorticotropic hormone; magenta). Scale bar: 50μm. d, qRT-PCR analysis revealing that the senescent marker Cdkn1a (encoding for p21Cip1) and the SASP components Il1b and Il6 are reduced in neoplastic pituitaries treated with 100 nM ouabain and 100 nM digoxin relative to vehicle controls (n = 3 per group). Statistical significance was calculated using unpaired two-tailed Student’s t-test; data represent mean ± s.d; n represents number of mice.
