Fig. 4.

Reduced expression level of polyadenylated crystallin mRNAs in rbm24a mutants. (A) RNA-seq analysis of whole embryos at 33 hpf. Volcano plot shows the down-regulation of lens-related genes coding for heat shock proteins and other crystallins in rbm24a mutants (represented by green dots). Red dots show up-regulated genes, with padj (P value adjusted) less than 0.05 and fold change more than 1.5; blue dots represent down-regulated genes, with padj less than 0.05 and fold change less than 0.67. (B) The qRT-PCR analysis compares crystallin mRNA levels in WT siblings and rbm24a mutants, following reverse transcription using oligo(dT) primer. The results were obtained from three independent experiments, using different batches of embryos. (C–L′) In situ hybridization analysis of crystallin gene expression patterns at 33 hpf. Representative images show strongly reduced expression of cryaa, hsp70, and hsp90aa1.2 in rbm24a mutants. The overall transcript abundance for other crystallin genes does not change. Due to the extremely high level of nucleotide sequence identity, the crygm2d1-21 probe recognizes transcripts of all 21 types of γ-crystallins. (Scale bar, 100 µm.)