human liver endothelial cells (hlecs)  (Lonza)

Journal: The Journal of Biological Chemistry

Article Title: Platelet Activation Receptor CLEC-2 Regulates Blood/Lymphatic Vessel Separation by Inhibiting Proliferation, Migration, and Tube Formation of Lymphatic Endothelial Cells *

doi: 10.1074/jbc.M111.329987

Figure Lengend Snippet: Inhibitory effects of platelets on endothelial cell proliferation. Platelets inhibited hLEC proliferation but not HUVEC proliferation, depending on CLEC-2. A, cell proliferation of hLECs (upper panels) and HUVECs (lower panels) in the presence of buffer (left panels) and hPlt (right panels, 1 × 108/ml) was investigated by thymidine analog incorporation assay. A group of EdU-incorporated cells is indicated by arrows. B and C, cell proliferation of hLECs (B) and HUVECs (C) in the presence of buffer, hPlt (1 × 108/ml), mPlt WT (1 × 108/ml), and CLEC-2-deficient murine washed platelets (mPlt KO, 1 × 108/ml) was investigated by thymidine analog incorporation assay. Quantification of the proliferation was performed as described under “Experimental Procedures.” The graph illustrates percent change ± S.E. from base line (buffer) (n = 10 from four independent experiments).

Article Snippet: Cells Human umbilical vein endothelial cells (HUVECs) and human lymphatic endothelial cells (hLECs) were purchased from Lonza (Basel, Switzerland) and maintained on culture dishes in endothelial growth medium-2 (EGM-2; Lonza) supplemented with 5% FBS and an EGM-2 microvascular set (0.5 ml of human EGF, 0.2 ml of hydrocortone, 25 ml of FBS, 0.5 ml of VEGF, 2 ml of human FGF-B, 0.5 ml of R3-IGF-1, 0.5 ml of ascorbic acid, and 0.5 ml of GA-1000).

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Platelet Activation Receptor CLEC-2 Regulates Blood/Lymphatic Vessel Separation by Inhibiting Proliferation, Migration, and Tube Formation of Lymphatic Endothelial Cells *

doi: 10.1074/jbc.M111.329987

Figure Lengend Snippet: Inhibitory effects of platelets on tube formation of endothelial cells through CLEC-2. A and B, tube formation of hLECs (4 × 105/ml) (A) or HUVECs (5 × 105/ml) (B) in the presence (hPlt) or absence (buffer) of human washed platelets (1 × 108/ml). Images are representative of five different experiments. C, quantification of tube formation. D, tube formation of mLECs (1.25 × 105/ml) in the presence of buffer, wild-type mPlt WT (1 × 107/ml), and CLEC-2-deficient murine washed platelets (mPlt KO, 1 × 107/ml). Images are representative of three different experiments. E, quantification of mLEC tube formation. The graphs in C and E show quantification of tube formation as percent change ± S.E. from base line (buffer) (n = 10–12 from three independent experiments). Three asterisks denote p < 0.005.

Article Snippet: Cells Human umbilical vein endothelial cells (HUVECs) and human lymphatic endothelial cells (hLECs) were purchased from Lonza (Basel, Switzerland) and maintained on culture dishes in endothelial growth medium-2 (EGM-2; Lonza) supplemented with 5% FBS and an EGM-2 microvascular set (0.5 ml of human EGF, 0.2 ml of hydrocortone, 25 ml of FBS, 0.5 ml of VEGF, 2 ml of human FGF-B, 0.5 ml of R3-IGF-1, 0.5 ml of ascorbic acid, and 0.5 ml of GA-1000).

Techniques: