Fig. 11.

Loss of RHOQ expression leads to NICD degradation in lysosomes. HUVECs cultured on BSA or rhDLL4 (1 μg/ml)-coated plates and effects on lysosomes were assessed 16 h later by staining cells with lysosome tracker (red) with changes in tracker levels, a visualised by confocal microscopy (nuclei stained with DAPI (blue)) and assessed by b FACs or cells were fixed and immuno-fluorescence staining for localisation changes in proteins c duo-link staining detecting only Lamp/Notch1 (red) or Lamp1/NICD (green) associated antibodies, visualised by confocal microscopy. Nuclei stained with DAPI (blue). d Transfected HUVECs with RHOQ siRNA duplexes were cultured on BSA or rhDLL4 (1 μg/ml)-coated plates for 8 h with or without chloroquine (10 µM) before (a) changes in DLL4/Notch downstream target expression by QPCR (Error bars = S.D. Key: *p < 0.05, **p < 0.01, ***p < 0.0001 one-way ANOVA or unpaired Student’s t-test between data group and control, Scale bar = 20 nm; representative images and data of n = 3 independent experiments)