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. 2014 Jun 1;127(11):2407–2419. doi: 10.1242/jcs.130344

Fig. 3.

Fig. 3.

NOTCH1 accumulates in late endosomes but is decreased in recycling endosomes. (A–H) hTERT-RPE1 cells immunostained with antibodies against endogenous NOTCH1 (green) and markers of late endosomes (RAB7, red) or multi-vesicular bodies (TSG101, red). Scale bars: 2 µm. (I,J) Quantification of the proportion of either RAB7-positive or extra-nuclear TSG101-positive endosomes that are also positive for NOTCH1 staining shown as a percentage of total RAB7- or TSG101-positive endosomes. Data show the mean±s.d. (K,L) hTERT-RPE1 cells coimmunostained for endogenous NOTCH1 (green) and recycling endosomes (RAB11, red). All immunofluorescence images represent a single plane of focus at the denoted endosomes and are at 100× magnification, with enlarged regions outlined by white boxes and enlarged individual vesicles outlined by dashed boxes and shown as insets. Scale bars: 2 µm (M) Quantification of the proportion of RAB11-positive endosomes that are also positive for NOTCH1 shown as a percentage of total RAB11-positive endosomes. Data show the mean±s.d. *P≤0.001 compared with control (Student's t-test).