Journal: Nature cell biology

Article Title: FBP1 loss disrupts liver metabolism and promotes tumourigenesis through a hepatic stellate cell senescence secretome

doi: 10.1038/s41556-020-0511-2

Figure Lengend Snippet: Decreased FBP1 expression in human and murine liver tumours (a) Metabolic gene set analysis of TCGA HCC RNA-sequencing data. A total of 374 HCC tumours and 50 adjacent normal tissues were included, and 2752 genes encoding all known human metabolic enzymes and transporters were classified according to the Kyoto Encyclopedia of Genes and Genomes (KEGG). Generated metabolic gene sets were ranked based on their median fold expression changes in HCC tumours vs normal tissue, and plotted as median ± median absolute deviation. (b) Representative FBP1 IHC staining on human liver tissue array with adjacent normal, grade 2 and 3 HCC tissues. Scale bar: 100 μm. ( c ) Statistical analysis of FBP1 IHC staining in ( b ). n=20 for normal, n=30 for grade 2, n=30 for grade 3 samples. In each box plot, the top-most line is the maximum, the top of the box is the third quartile, the centre line is the median, the bottom of the box is the first quartile and the bottom-most line is the minimum. ( d ) Representative H E staining in liver sections from 24-week control (Ctrl) and DEN-treated (DEN) C57BL/6 mice (n = 3 independent experiments with similar results). T, tumour. Scale bar: 100 μm. (e) Serum alanine transaminase (ALT) activity from 24-week Ctrl and DEN mice. n=4 for Ctrl, n=5 for DEN. Graph in e show mean ± SEM, and P value was calculated using a two-tailed t-test. Numerical source data are provided in Source Data Extended Data Fig. 1.

Article Snippet: Primary mouse HSCs were isolated from 10-week C57BL/6 mice following a previous protocol .

Techniques: Expressing, RNA Sequencing Assay, Generated, Immunohistochemistry, Staining, Mouse Assay, Activity Assay, Two Tailed Test

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 × 10 5 /ml) ( A ) or HUVECs (5 × 10 5 /ml) ( B ) in the presence (hPlt) or absence (buffer) of human washed platelets (1 × 10 8 /ml). Images are representative of five different experiments. C , quantification of tube formation. D , tube formation of mLECs (1.25 × 10 5 /ml) in the presence of buffer, wild-type mPlt WT (1 × 10 7 /ml), and CLEC-2-deficient murine washed platelets ( mPlt KO , 1 × 10 7 /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

Article Snippet: 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 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 × 10 8 /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 × 10 8 /ml), mPlt WT (1 × 10 8 /ml), and CLEC-2-deficient murine washed platelets ( mPlt KO , 1 × 10 8 /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: 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: