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(A) Wild-type zebrafish embryo axial musculature is highly birefringent at 72 hpf. (B) APM [6 µM] exposure from 12–72 hpf resulted in defects in the axial musculature, which are shown by a reduced birefringence at 72 hpf.

APM exposures for (A) 24 h (26–50 hpf) and (B) 2 h (48–50 hpf). Concentrations are given in µM. Bars represent the relative gene expression as fold change of the respective untreated control as mean ± standard deviation of three independent replicate exposures. Control  =  ctrl. * P<0.05.

(A) hspb11 expression analysis in ache and sop^fixe−/− zebrafish mutant embryos at 48 hpf. (B) hspb11 expression in sop^fixe null mutants and siblings (+/?  =  heterozygous and wildtype) after APM (6 µM) exposure (26–50 hpf). (C) 1 µM Thapsigargin (TG) and 2 mM caffeine (CAF) induce hspb11 expression in zebrafish embryos (exposure period 48–50 hpf). Bars represent the relative gene expression as fold change of the respective untreated control as mean ± standard deviation of three independent replicate exposures Control  =  ctrl. * P<0.05.

Expression pattern of hspb11 at 18 hpf were detected in developing adaxial musculature (A–A″). During later developmental stages, at 24 hpf, hspb11 mRNA transcripts were restricted to muscle pioneers (B and B′) and became only expressed in the notochord at 50 hpf (C). (A, B) lateral view, (A′) dorsal view and (A″ and B′) cross sections. Increased expression pattern in axial musculature was observed in APM exposed (D) and homozygous (−/−) ache mutant embryos (E). Hspb11 expression in homozygous sop^fixe−/− embryos (F and G) and heterozygous/wildtype (+/?) siblings (H and I).

(A) qPCR hspb11 expression analysis. (B) Wildtype hspb11 expression pattern at 24 hpf. (C) MS222 treatment results in the loss of hspb11 expression pattern at 24 hpf.

(A, D and G) mmMO-hspb11 injected embryos. (B, E and H) MO(ATG)-hspb11 morphants. (C, F and I) MO(UTR)-hspb11 morphants. (A–C) Phenotypic observations of morpholino injected embryos at 48 hpf. (D–E) Muscle organization determined with birefringence at 72 hpf. (G–I) Slow muscle myosin distribution at 72 hpf shown by immunofluorescence staining with antibody F59. Arrows indicate gaps in fiber distribution.