Figure 1

Establishment of the hsf4 knockout zebrafish. (a) All 13 exons of zebrafish hsf4 and the TALEN target site were shown. The TALE sequences were highlighted in red. (b) Bsr1 assay. The upper picture was the genotyping results. PCR products from F2 zebrafish and wild type were subjected to the Bsr1 digestion. The wild-type allele cannot be digested, thus, only had one 466-bp band. The del7 allele could be digested by Bsr1, producing two bands that were 305 and 161 bp, respectively. Lanes (from left to right): three samples, WT control, negative control and size standard. The lower sequence showed the new Bsr1 restriction site generated by the 7-bp deletion in the mutant fish. (c) Sanger sequencing of WT and hsf4 mutant fish (del7, c.211-217del). The 7-bp deletion was pointed out with a box. (d) Western blot analysis of the hsf4 protein in the WT and hsf4^null zebrafish. The zebrafish hsf4 protein expressed in eukaryotic cells was used as a positive control. Tubulin was a loading control and the ov-hsf4 sample from the HLECs was the positive control. The anti-hsf4 antibody could recognize the translated product of the transcript XM_009293553(424aa). The black arrow pointed to the hsf4 band