Fig. 5

Notch signaling regulates Rab10 GTPase activity though DENNd4C. (a) Graph of relative gene expression of DENNd4A,B,C in ECs cultured in VEGF-containing or serum-starve (SS) media. Gene expression levels normalized to GAPDH. N number of replicates. (b) Representative images of indicated siRNA-treated ECs stained for collagen IV (Col IV) (red), actin (green), and DNA (blue). Dotted green lines indicates cell outline. Arrowheads denote extracellular Col IV secretion. (c) Graph of collagen IV (Col IV) extracellular ratio in indicated siRNA-treated ECs. (d) Representative images of indicated siRNA-treated ECs stained for Col IV (red), LH3 (green), actin (light blue), and DNA (blue). Yellow arrowheads indicate co-localized puncta only, red arrowheads indicate Col IV only puncta, and green arrowheads indicate LH3 puncta only. (e) Graph of percent CIVCs containing LH3 in indicated siRNA-treated ECs. (f) Representative images of ECs expressing DENNd4C-flag and stained for Col IV (green) and LH3 (red). Yellow arrowheads indicate Col IV-containing vesicles co-localized with LH3 and red arrowheads indicate LH3 co-localized with DENNd4C-flag. (g) Graph of percent Col IV-containing vesicles with LH3 in ECs expressing DENNd4C-flag in indicated culture media. For all graphs n number of cells unless otherwise indicated. Black bars indicate comparison groups with indicated p-values. All p-values are from two-tailed Student’s t-test from at least three experiments. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.0001; ns not significant