Fig. 4

Zebrafish Hsp60 knockout results in early larval developmental abnormalities. (A) Hsp60 protein levels of hspd1+/+, hspd1+/−, and hspd1−/− larvae at 3 dpf, and hspd1−/− larvae at 3, 5, and 7 dpf. Assessed by western blot (n = 6). (B) Lateral imaging of hspd1−/− larvae at 3, 5, and 7 dpf indicated developmental delay at 5 and 7 dpf, but hspd1+/− larvae showed no significant phenotypic differences compared to hspd1+/+ larvae. (C) Quantification of WT and KO hspd1 transcript levels in hspd1+/+ and hspd1−/− larvae at 5 dpf, quantified by RNA sequencing. (D–F) Phenotypic quantification of body length (D), eye size (E), and swim bladder inflation (F) in hspd1+/+ (n = 28), hspd1−/− (n = 31) larvae at 3, 5, and 7 dpf. (G–H) Volcano plots showing analysis of transcriptome (G), and proteome (H) changes between hspd1+/+ and hspd1−/− larvae. Cut-off values are indicated with dashed lines. Mitochondrial proteins based on MitoCarta3.0 (MC3.0; [ 2 ]) are highlighted with red points. (I) Comparison plot of log2fold changes of Proteomics versus Transcriptomics in hspd1−/− larvae. Cut-off values are indicated with dashed lines. Mitochondrial proteins are highlighted in red. Genes significantly up- or down-regulated at both transcript and protein level are labeled. (J) Violin plots distinguishing proteins from mitochondrial sub-compartments of hspd1−/− larvae in comparison to hspd1+/+ larvae. Statistical significance was determined with one-way ANOVA: ∗p < 0.05, ∗∗∗∗p < 0.0001. Abbreviations: dpf, days post-fertilization; KO, knockout; sgRNA, single guide RNAs; WT, wild type.