Figure 1. Pericentrin (pcnt) mRNA is localized to centrosomes in early zebrafish embryos.

(A) RNA in situ hybridization of transcripts of different PCM components in four-cell stage zebrafish embryos. Note that while the mRNA of cep152, cep192, and cep215 displayed a pan-cellular distribution, pcnt mRNA was concentrated at two distinct foci in each cell. (B) RNA in situ hybridization showed similar dot-like patterns of pcnt transcripts with two non-overlapping antisense probes. The signals were lost in two maternal-zygotic (MZ) pcnt mutants. (C) Fluorescent RNA in situ hybridization and anti-γ-tubulin co-staining demonstrated the centrosomal localization of pcnt mRNA. n > 300 (pcnt-001 probe), n > 100 (pcnt-002 probe), n > 50 (cep152, cep192, or cep215 probe); all the embryos showed the same RNA distribution patterns as shown in the representative images. More than 100 MZpcnt^tup2 or MZpcnt^tup5 embryos were examined; none of them showed visible pcnt RNA in situ signals. Embryos were examined between 2- and 16-cell stages with representative four-cell stage embryos shown. Dashed lines delineate the cell boundaries. Scale bars: 200 µm or 25 µm (inset in C).

Figure 1—figure supplement 1. Sequences of two Cas9-induced frameshift mutations (alleles pcnt^tup2 and pcnt^tup5) in the zebrafish pcnt gene.

The wild-type reference sequence is on the top. The guide RNA targets the exon 2 of the pcnt transcript (encoded by ENSDARG00000033012). The target site is underlined and the protospacer-adjacent motif (PAM) is in orange (on the reverse strand). Insertions and deletions (indels) are indicated by blue lowercase letters and dashes, respectively. The net change of each indel mutation is noted at the right of each sequence (+, insertion; −, deletion).