Figure 2.

Partial antisense repression of R-Spondin1 restores the regenerative ability in the presence of TCDD. A) The bold line at the intron-exon (I1-E2) boundary indicates the gene sequence targeted with the R-Spondin1 splice variant MO. To analyze the splice blocking efficiency, forward and reverse primers (arrows at E1 and E3) were designed for RT-PCR. B) The fin tissue of control and zfR-Spondin1 morphants were amputated at 2 dpf and exposed to vehicle or TCDD. The fin regeneration images were taken at 3 dpa. The dotted line marks the plane of amputation. The experiments were repeated multiple times, and ∼80% of the R-Spondin1 morphants did not elicit inhibition of regeneration after TCDD exposure (n=70/85). C) Analysis of R-Spondin1 transcript in control and R-Spondin1 morphants (3 dpf) after qRT-PCR, followed by agarose gel electrophoresis (2%). The arrow points to the mispliced R-Spondin1 variant after the loss of the targeted exon2. β-Actin expression was used as the loading control.