Fig. 6

The expression of ntng2b was analyzed and overexpression promoted M-cells axon regeneration. (A) Quantitative histogram of netrin family expression analysis by RNA-seq between experiment and control group. Assessed by unpaired, two-tailed Student's t-test. * p < 0.05, ns, no significance. (B) Line graph of relative expression analysis of ntng2b in zebrafish larvae by qRT-PCR at different developmental stages. The total RNA of each stage was obtained from 30 zebrafish larvae by three independent experiments. (C) Histogram of relative expression analysis of ntng2b by qRT-PCR in adult zebrafish at different tissues. The total RNA of each tissue was obtained from 4 adult zebrafish by three independent experiments. (D) Statistical quantitative diagram of axon regeneration (control: 254.9 ± 30.71 μm, n = 7; Ntng2b overexpression: 484.8 ± 47.62 μm, n = 7). Assessed by unpaired, two-tailed Student's t-test. ** p < 0.01. (E) Representative diagram of confocal imaging of M-cells regenerated axons and branches. Asterisk, ablation site; Arrowhead, axon branch points; Scale bar, 50 μm. (F) Statistical quantitative diagram of axon branch point number (control: 1.57 ± 0.48, n = 7, total branch points number = 11; Ntng2b overexpression: 3.43 ± 0.48, n = 7, total branch points number = 24). Assessed by unpaired, two-tailed Student's t-test. * p < 0.05. (G) Timeline of time points of electroporation, axotomy and imaging. (H) Representative diagram of confocal imaging of M-cells regenerated axons. Asterisk, ablation site; Scale bar, 50 μm. (I) Statistical quantitative diagram of axon regeneration (Rab11 overexpression: 435.7 ± 28.23 μm, n = 7, Rab11bb overexpression / Ntng2b knockdown: 166.6 ± 25.97 μm, n = 6, Ntng2b knockdown: 139.1 ± 22.25 μm, n = 7). Assessed by unpaired, two-tailed Student's t-test. **** p < 0.0001, ns, no significance.