Wild-type (WT), zygotic pnrc2oz22
(Zpnrc2), maternal pnrc2oz22
(Mpnrc2), and maternal-zygotic
pnrc2oz22 (MZpnrc2) mutant
embryos were raised to mid-segmentation stage (16–18 hpf) and probed for
her1 (A–D) and dlc expression
(E–F) by in situ hybridization (n ≥ 7 each). WT,
Mpnrc2 mutant, and MZpnrc2 mutant embryos
(n = 10 per biological replicate) were analyzed by qPCR using primers to amplify
across exon-exon boundaries to detect spliced her1 (I) and
dlc (J) transcripts or primers to amplify across
intron-exon boundaries to detect her1 (K) and
dlc (L) unspliced transcripts. MZpnrc2
mutant embryos have ~4-fold higher levels of spliced
her1 and dlc mRNA than wild-type or
Mpnrc2 mutant embryos, which have comparable levels (I and
J). Both MZpnrc2 and Mpnrc2 mutant embryos
have ~2-fold less unspliced her1 and
dlc transcripts compared to WT embryos (K and L). hpf =
hours post-fertilization.