ZFIN Image: Tuschl et al., 2016, Fig. 5

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Figure Caption

Fig. 5

Zebrafish slc39a14 is expressed during early zebrafish development and significantly reduced in slc39a14^U801 mutants. (a) RT-PCR showing slc39a14 expression between 3 and 120 hpf in zebrafish. L, 100 bp ladder (Promega). (b) Whole-mount in situ hybridization using a DIG-labelled antisense RNA probe showing slc39a14 expression in the proximal convoluted (black arrows) and straight (red arrows) pronephric tubules in zebrafish larvae at 4 dpf. Top, lateral view; bottom, dorsal view. Scale bar, 200 µm. (c) DNA sequence of the region within exon 5 of slc39a14 targeted by a CRISPR guide RNA is highlighted in yellow and the 2-bp deletion introduced in the slc39a14^U801 mutant indicated by dashes. Pam sequence underlined. (d) qRT-PCR demonstrates a 2.2-fold reduction in slc39a14 expression in homozygous slc39a14^U801 mutants (*P=0.0117). Primers were designed to detect all slc39a14 transcripts (Supplementary Table 4). Ef1α was used as a reference gene. Data are presented as means±s.d. from three independent experiments. Statistical analysis was performed using Student’s two-tailed t-test on individual ΔCt values (*P<00.5).

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Acknowledgments

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