Expression of ZNF24 induces vascular defects in zebrafish embryos. Tg (Fli1:EGFP) embryos were injected with control (A-Aii) or ZNF24 mRNA (B-Bii, and images were taken at 30 hpf. (C) Embryos were injected with control or ZNF24 mRNA, and real-time PCR was performed using primers specific for VEGF and ELF1α. *P = .03. (D-Gii) Effects of ZNF24 expression can be rescued by overexpression of VEGF. Tg (Fli1:EGFP) embryos were injected with control (D-Dii), ZNF24 mRNA (E-Eii), ZNF24 mRNA and VEGF mRNA 25 ng/μL (F-Fii), or ZNF24 mRNA and VEGF mRNA 200 ng/μL (G-Gii), and images were taken at 30 hpf. Black asterisk indicates pericardial edema; white asterisk, abnormal caudal vascular plexus. (H) Percentages of defective embryos injected with indicated reagents. *P = .004 comparing embryos injected with ZNF24 mRNA alone and embryos injected with ZNF24 mRNA and VEGF mRNA 200 ng/μL. N indicates the number of experimental repeats; n the total number of embryos injected. Control: N = 3, n = 119; ZNF24: N = 3, n = 140; ZNF24 + VEGF 25 ng/μL: N = 3, n = 80; ZNF24 + VEGF 200 ng/μL: N = 3, n = 164. (I) Percentages of defective embryos injected with indicated reagents. *P = 8.09E-04 comparing embryos injected with ZNF24 mRNA alone and embryos injected with ZNF24 mRNA and VEGF promoter ODN. N indicates the number of experimental repeats; n the total number of embryos injected. Control: N = 3, n = 295; ZNF24: N = 3, n = 158; ZNF24 + Scrambled ODN: N = 3, n = 178; ZNF24 + VEGF ODN: N = 3, n = 147. Brightfield images were obtained at room temperature using a Nikon Eclipse 80i microscope equipped with a 4×/0.13 objective (Nikon), SPOT 7.4 Slider camera and SPOT software (Version 4.6, Diagnostic Instruments). Fluorescent images were obtained at room temperature using a Nicon Eclipse 80i microscope equipped with 4×/0.13 and 10×/0.30 objectives (Nikon), Retiga-2000R camera (QImaging), and NIS-Elements AR 3.0 software (Nikon).