Figure 5

Bmper was involved in endoglin-regulated vasculogenesis

(A) Heat maps presenting part of the RNA-Seq result. Six vasculogenesis genes were significantly decreased after endoglin knockdown, including bmper, kmt2e, use1, csnk2a1, tmem199 and sin3aa. The level of expression of the gene is indicated with red (high) or blue (low). (B) qPCR analysis of bmper expression in different stages of the Con-MO and eng-MO group, including 64 cell, sphere, 30% epiboly, 75% epiboly, 11 hpf and 24 hpf stage. Gapdh was used as an internal control (mean ± SD; the experiments were repeated three times). (C) Bmper expression by WISH in 24 hpf zebrafish embryos of the Con-MO and eng-MO group. The red arrow indicates the region where the expression of bmper was significantly decreased. (D) Fluorescence image of Con-MO, Con-MO + bmper mRNA, eng-MO and eng-MO + bmper mRNA group. (All Tg(fli1a:EGFP)^y1 zebrafish embryos were injected with 2 ng morpholinos and 500 pg mRNA. For example, Con-MO group: 2 ng 5-mispair control MO and 500 pg mCherry mRNA.) The red and white arrows point to the disrupted and rescued structure of DLAV and ISVs, respectively. (E) qPCR analysis for cdh5, dll4, flt4 and kdrl expression in fli-GFP⁺ cells of the Con-MO, eng-MO and eng-MO + bmper mRNA group. Gapdh was used as an internal control (mean ± SD; the experiments were repeated three times). A value of P was considered statistically significant (^*P<0.05) for B and E. Scale bars are 100 μm for C and D.