Fig. 7. EGFL7 is associated with the interstitial ECM. (A) Anti-HA western blot on different fractions of chicken embryonic fibroblasts expressing GFP or HA-tagged human EGFL7 (top panel), and Coomassie Blue staining of the same cell fractions prepared from an equal number of cells (bottom panel). (B-G) The following cells were cultured for 3 days and then double stained for EGFL7 (green) and with DAPI (blue) in the absence of detergent: HUVECs (dark blue ovals in diagram) and human dermal fibroblasts (light blue ovals) in mixed culture (B); HUVECs alone (C); fibroblasts alone (E); HUVECs in the bottom well with fibroblasts in an insert (D); or HUVEC alone on fibronectin (F) or laminin (G). (H-M) Confocal images of aortic ring sprouts from Egfl7^+/+ (H-K) or Egfl7^-/- (L,M) littermates stained for fibronectin (blue), EGFL7 (red), CD31 (green) and with DAPI (gray). H-J show a segment of sprout with stalk cells; K shows tip cells; L,M show a segment of sprouts with both tip and stalk cells. The actual size represented by the width of the panel: 650 µm (B-G); 80 µm (H-J); 54 µm (K); 100 µm (L,M).