Figure 7.

a2V ablation impairs early endosomal trafficking resulting in an increase of TGF-β signaling. Inguinal mammary glands from virgin WT and KO mice were evaluated for the status of TGF-β and its downstream effectors. (a) Localization of TGF-β (green) and early endosome marker EEA1 (red) in WT and KO mice by immunofluorescence. Nucleus was stained with DAPI (diamidino-2-phenylindole; blue). Scale bars: 10 μm. (b) Fold increase in mRNA expression levels of Notch target gene Hey1, and TGF mediator Smad2 were assessed by quantitative real-time-PCR (qRT-PCR). Prior to fold-change calculation, the values were normalized to endogenous control glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Data represent mean±standard error, n=6. ***P≤0.001. (c) Confocal microscopy for nuclear localization of pSmad2 (green) in pubertal mice. Nucleus was stained with DAPI (blue). Scale bars: 20 μm