Extended Data Fig. 3: Impact of CDK4/6 inhibition and hypoxia on CCL2 secretion.
a, CCL2 secretion from TS/A cells exposed to palbociclib (P), tamoxifen (T) or their combination, for 72 h (n = 8 from two independent experiments for control, n = 5 from two independent experiments for T, n = 6 from two independent experiments for P and P + T). Box-plots overlaid by individual data points show medians ± interquartile range; whiskers indicate minima and maxima. Relevant and significant (P < 0.05) statistical differences (by linear mixed-effects regression followed by simultaneous tests of general linear hypotheses) are shown. b, Workflow followed for the quantification of pimonidazole positive area in M/D-driven tumor tissues. Tiled z-stack images were analyzed with Image J. Nuclei and pimonidazole staining images were individually processed with z-projection (sum all slices) and followed by adjusting the threshold in the Otsu method. Regions of interest (ROIs) were selected by auto-detection or manually based on nuclear staining and applied to the pimonidazole staining image to measure pimonidazole-positive area. For representative images and quantification of pimonidazole-positive area, see Fig. 3i.
