Extended Data Fig. 3: Age-associated protein expression in breast cancer.

Immunohistochemistry (IHC) of tissue microarrays (TMAs) from a, Vancouver Big series, and b, METABRIC series. Even though pathologists score TMA staining patterns with a variety of schemas, e.g. percent of tumour cells positively stained, staining intensity, or a combination thereof, biphasic and other age-associated patterns are still revealed via supersmoother fits to the data. The age at which biphasic inflection occurs varies by biomarker, e.g. near 60 years of age for EZH2 and p16, near 70 years for ER. The red curves are supersmoother curves fitted to the IHC data. The tan coloured intervals are randomization test confidence bands under the null hypothesis of no age association (see Methods) at one of the four confidence levels indicated by the associated p-value bound in each graph. Red curves that extend beyond the (1-α) confidence bounds at any location indicate age-association at the given confidence level with a two-sided p-value p < α (95%, 99%, 99.9% and 99.99% confidence levels assessed via randomization tests yielding p < 0.05, 0.01, 0.001 and 1e-04 respectively, exact p-values not available, see Methods).