Extended Data Fig. 4: Loss of tango2 leads to reduced locomotor activity of zebrafish larvae.

a, Strategy to knock out tango2 in zebrafish and verification of the knockout by PCR genotyping. The sgRNA targeting sites and the positions of PCR primers are indicated. b, Comparison of the sequences between the wild type tango2 and the mutated allele in zebrafish. The exon2 - intron2 junction is deleted in the mutant allele. c, Verification of intron 2 retention in the tango2^−/− zebrafish at the mRNA level by reverse transcription (RT)-PCR. d, The predicted protein product of Tango2 in tango2^−/− zebrafish. e, Whole-mount in situ hybridization showing maternal expression of zebrafish tango2. Arrowheads indicate the expression of tango2 in 1-cell and 1000-cell stage embryos. Arrows indicate the expression of gata1 (control) in the intermediate cell mass and tango2 in neuronal tissues at 24 h post-fertilization (hpf). Scale bars, 200 μm. f, o-dianisidine staining of wild type and tango2-knockout zebrafish embryos at 48 and 72 hpf. Scale bars, 200 μm. g, Total haem content in wild type and tango2-knockout zebrafish embryos at 48 and 72 hpf. n = 3 independent experiments. Data are presented as mean ± s.e.m. h, Whole-mount in situ hybridization showing normal expression of the erythroid markers gata1 and αe3-globin in tango2-knockout zebrafish embryos. Scale bars, 200 μm. i, Knockout of tango2 did not reduce erythrocytes in Tg(globin-LCR:EGFP) transgenic zebrafish embryos. n = 3 independent experiments. Data are presented as mean ± s.e.m. j, Distance travelled per minute by 12-dpf wild type and tango2-knockout zebrafish larvae during the 10-min dark / 10-min light cycles. Black bars indicate dark and white bars indicate light. n = 16 fish for each group. k, Representative swimming tracks of 12-dpf wild type and tango2-knockout zebrafish larvae over a 40-min period under constant light. Scale bars, 2.5 mm. Statistical significance was determined by one-way ANOVA followed by Tukey’s test.

Source data