Fig. 1

Over-expression of human DLG5 causes thrombocytes loss in zebrafish. (a–h) Gross morphology of Tg (CD41: GFP) zebrafish embryos at 3-dpf. Compared with control zebrafish (empty vector), human DLG5 over-expression (DLG5+) causes pericardial oedema (e, red arrowhead) and thrombocyte loss (g, h, yellow arrows). (i,j) Quantification of the average number of thrombocyte (GFP+) at caudal vein. Error bars, mean ± SEM.; *P < 0.05 (n = 10; Student's t test). dpf, days post fertilization.

Fig. 2

Human DLG5 over-expression prevent phenylhydrazine (PHZ)-induced zebrafish thrombosis. (a–l) Representative images of Tg (CD41: GFP) zebrafish embryos at 3-dpf treated with WT control (empty vector), human DLG5, WT control plus PHZ (2 μM) or human DLG5 plus PHZ (2 μM). Zebrafish at 2-dpf treated with 2-μM PHZ for 24 h visually showed severe thrombosis and platelet aggregation in the caudal vessels (c,g,k, red arrows) without observable toxicity. (c,d,g,h) Compared with PHZ treated group, human DLG5 over-expression prevent PHZ-induced zebrafish thrombosis. (m) Quantification of the average number of thrombocyte (GFP+) at caudal vein. Columns, mean; bars, SEM (n = 10; ANOVA; * P < 0.05). dpf, days post fertilization; PHZ, phenylhydrazine.

Fig. 3

Effect of human DLG5 over-expression on signalling pathways in zebrafish. (a) Endogenous mpl, itga2b (ITGA2B), itgb3b (ITGB3), tubb1, wasb (WAS), actn1, myh9a (MYH9), gp1bb, gp9, flna, prkacaa, vwf, nbeal2, cycsa (CYCS), abcg5, abcg8, dlg5a (DLG5), mst1, stk3 (MST2), lats1, lats2, yap1 (YAP) and taz in WT control (empty vector) and human DLG5 injected embryos assessed by qRT-PCR (n = 10 individual embryos). * P < 0.05. (b) DLG5 over-expression causes changes in expression levels of ‘genes’ involved in MK growth, differentiation and maturation. The red and blue arrows colours indicate upregulated and downregulated molecules, respectively.

Fig. 4

Effect of human DLG5 over-expression on signalling pathways in megakaryoblast leukaemia (MEG)-01 cells. (a–g) DLG5, LAST1, LAST2, MST2, YAP and TAZ in control and DLG5-IRES-EGFP plasmids were transfected into MEG-01 cells assessed by qRT-PCR. * P < 0.05.

Acknowledgments
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