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:

Journal: PLoS ONE

Article Title: High Mobility Group Box-1 Promotes Inflammation-Induced Lymphangiogenesis via Toll-Like Receptor 4-Dependent Signalling Pathway

doi: 10.1371/journal.pone.0154187

Figure Lengend Snippet: (A): HMGB1 promoted VEGF-C-induced HDLECs proliferation in a dose-dependent manner. (B): TLR4 mediates HMGB1-induced LECs proliferation. (C-E): TLR4 mediates HMGB1-induced LECs tube formation.* p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Human dermal lymphatic endothelial cells (HDLECs) were purchased from ScienCell (Carlsbad, CA) and maintained in endothelial cell basal medium-2 with growth supplements (EBM-2 MV).

Techniques:

Journal: Respiratory Research

Article Title: Apoptosis of viral-infected airway epithelial cells limit viral production and is altered by corticosteroid exposure

doi: 10.1186/1465-9921-7-78

Figure Lengend Snippet: Representative images of airway epithelial immunostaining of Cam Hartley Guinea pigs Semi-quantitative scoring was utilized to determine the expression for DR4(A), DR5 (C) and p85-PARP (E) in immunohistochemically stained lung sections. Panels B, D and F were the isotype controls for the respective antibodies. Arrows indicate the stained epithelial cells. Scale bar represent 10 μm in panels A through F.

Article Snippet: Polyclonal rabbit anti-DR4 and -DR5 antibodies (Cell Sciences Inc, MA) and rabbit anti- PARP p85 fragment antibody (Promega, MA) were used along with normal Rabbit IgG as negative control to measure receptor expression and apoptosis respectively. p85-PARP antibody is specific for the p85 fragment of PARP generated by caspase cleavage and provides a reliable measure of in situ apoptosis [ ].

Techniques: Immunostaining, Expressing, Staining

Journal: Respiratory Research

Article Title: Apoptosis of viral-infected airway epithelial cells limit viral production and is altered by corticosteroid exposure

doi: 10.1186/1465-9921-7-78

Figure Lengend Snippet: Acutely infected GPTEC demonstrate apoptosis coordinate with DR4 and DR5 expression Semi-quantitative scoring was utilized to determine the expression of p85-PARP, DR4 and DR5 in the Guinea pig lung sections by immunohistochemistry. p85-PARP was significantly higher in 1 -4 dPi lung sections compared to Sham controls, peaked at 4 dPi and decreasing significantly by 7 dPi (Figure 4A). This trend in apoptosis in the Acute model was coordinate with the changes in DR4 and DR5 expression (Figure 4B). * p < 0.05 compared to Sham and †p < 0.05 compared to 4 dPi.

Article Snippet: Polyclonal rabbit anti-DR4 and -DR5 antibodies (Cell Sciences Inc, MA) and rabbit anti- PARP p85 fragment antibody (Promega, MA) were used along with normal Rabbit IgG as negative control to measure receptor expression and apoptosis respectively. p85-PARP antibody is specific for the p85 fragment of PARP generated by caspase cleavage and provides a reliable measure of in situ apoptosis [ ].

Techniques: Infection, Expressing, Immunohistochemistry

Journal: Respiratory Research

Article Title: Apoptosis of viral-infected airway epithelial cells limit viral production and is altered by corticosteroid exposure

doi: 10.1186/1465-9921-7-78

Figure Lengend Snippet: Guinea pig AEC apoptosis and DR expression as detected by Immunohistochemistry of the Chronic model of Guinea pig viral infection and airway inflammation Semi-quantitative scoring was utilized to determine the expression of p85-PARP, DR4 and DR5 in the Guinea pig lung sections by immunohistochemistry. Significant reduction in the detection of p85-PARP for Ad5+OVA+Bud group (** p < 0.005) was observed when compared to Ad5 alone group (Figure 7A). However, DR4/DR5 expression for Ad5+OVA+Bud group was higher compared to Ad5 alone group * p < 0.05 (Figure 7B).

Article Snippet: Polyclonal rabbit anti-DR4 and -DR5 antibodies (Cell Sciences Inc, MA) and rabbit anti- PARP p85 fragment antibody (Promega, MA) were used along with normal Rabbit IgG as negative control to measure receptor expression and apoptosis respectively. p85-PARP antibody is specific for the p85 fragment of PARP generated by caspase cleavage and provides a reliable measure of in situ apoptosis [ ].

Techniques: Expressing, Immunohistochemistry, Infection