Fig. 3: Pipp ablation increases the size of Pten+/− mammary organoids.
A, B Mammary organoids derived from wild-type, Pipp−/−, Pten+/− and Pipp−/−;Pten+/− mice were cultured in growth factor reduced Matrigel for 14 d and imaged by brightfield microscopy (A). Data represent the mean organoid size ± SEM relative to wild-type organoids which were arbitrarily assigned a value of 1 (B) (n = 3 independent experiments, >87 organoids/genotype/experiment). C FFPE sections of mammary organoids derived from wild-type, Pipp−/−, Pten+/− and Pipp−/−;Pten+/− mice cultured in Matrigel were stained with Alexa-Fluor 488 phalloidin and DAPI. D, E Mammary organoids derived from wild-type, Pipp−/−, Pten+/− and Pipp−/−;Pten+/− mice were cultured in growth factor-reduced Matrigel for 14 d and imaged by brightfield microscopy (D). Data represent the mean percentage of branched organoids ± SEM (E) (n = 4 independent experiments, 138-623 organoids/genotype/experiment). F FFPE sections of mammary organoids derived from wild-type, Pipp−/−, Pten+/− and Pipp−/−;Pten+/− mice cultured in Matrigel were immunostained with cytokeratin 8 and cytokeratin 14 antibodies. G, H FFPE sections of mammary organoids from wild-type, Pipp−/−, Pten+/− and Pipp−/−;Pten+/− mice were immunostained with a pS6 antibody (G). Data represent mean pS6 fluorescence intensity ± SEM (H) (n = 4 independent experiments, >138 organoids/genotype/experiment). Scale bars, 500 μm (A), 100 μm (C, D, F, G).
