Figure 1. Morpholino (MO) knockdown of foxk1/foxk2a/foxk2b and transcriptional profiling of triple foxk morphants
A) Upper panels:Zebrafish larvae (72 hpf) injected with morpholinos targeting single foxk transcripts. Lower panels: triple (foxk1/foxk2a/foxk2b) MO and rescue with full length (+ hfoxk1 RNA) or mutated (+ hfoxk1-mut) human foxk1 transcripts, at 24hpf. Lower right panel: Hierarchical clustering of embryo numbers corresponding to: triple MO, triple MO with full length foxk1 transcript, triple MO with mutated foxk1 transcript, and wild type, divided into phenotype severity groups (weak/wt: no notable developmental delay; mild: minor developmental delay, trunk curvature, pigmentation issues; strong: severe developmental delay, nervous system defects, shortened body axes). B) Similarity distance matrix of RNA-seq data (counts) corresponding to wild type (wt) and triple morpholino (MO) conditions. C) MA plot displaying upregulated and downregulated genes (red dots) caused by triple Foxk MO. D) Expression profiles (TPM) of previously described Foxk targets (fbxo32, ulk1b) in wt and triple MO conditions. E) Gene ontology analysis of the most significantly (n=500) upregulated and downregulated genes in the triple MO condition.