Knockdown of larp7 and bcdin3 in zebrafish caused developmental defects, decreased 7SK levels, and aberrant splicing. (A and B) RNA in situ hybridizations of whole-mount zebrafish embryos by using larp7 and bcdin3 antisense (1, 2) and sense probes (3, 4) at the indicated time points. Black arrowheads in panels 1 and 2 indicate larp7 and bcdin3 expressions in blastomeres and their enriched expressions in developing brain/head, respectively. (C) Zebrafish embryos injected with either larp7 MO or bcdin3 MO show developmental defects at 24 hpf compared with embryo injected with a control MO. Black arrowheads indicate the developing brain/head and trunk/tail. (D) Relative quantities of 7SK levels were determined by RT-qPCR at the indicated time points below the graph. The error bars represent the mean ± SD. (E) Splicing pattern analysis for the 2 genes indicated on the left at different time points as shown above the agarose gels. Levels of splicing variants were determined by RT-PCR by using the samples as in D and gene-specific primers.
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