Figure 2.
Injections of mutated mRNAs into srp54+/− embryos induce an SDS-like phenotype, but the residual neutrophils are sufficient to be adequately recruited to injury sites. (A-D) Injection of T115A, T117Δ, or G226E human mRNA into srp54+/− embryos. (A-B) Representative images with (C-D) corresponding quantifications after WISH for mpo (A,C) and trypsin (B,D). (E) Fluorescent confocal microscopy images 8 hours after tail fin injury. Left, bright field; middle, fluorescence; right, merge. Yellow rectangle indicates the analyzed tail region. (F-H) Quantification of neutrophil migration of WT siblings, WT siblings injected with human G226E mRNA, srp54+/−, and srp54+/− zebrafish injected with human G226E mRNA. (F) Total number of mpo+ cells, and (G) mpo+ cells at wound. (H) Fraction of cells migrating toward the injury site. Images were acquired with a Point Scanning Confocal Leica SP5-II-MATRIX microscope (original magnification, ×10). Neutrophils were automatically counted using ImageJ software (we used 3 biological replicates with at least 2 to 3 larvae per replicate). Student t test was used for statistical analysis. ns, not significant. *P < .05; **P < .01; ***P < .005. Bar plots and horizontal lines in the graph represent the mean value of the replicates. Error bars indicate the standard deviation of the mean.

Injections of mutated mRNAs into srp54+/− embryos induce an SDS-like phenotype, but the residual neutrophils are sufficient to be adequately recruited to injury sites. (A-D) Injection of T115A, T117Δ, or G226E human mRNA into srp54+/− embryos. (A-B) Representative images with (C-D) corresponding quantifications after WISH for mpo (A,C) and trypsin (B,D). (E) Fluorescent confocal microscopy images 8 hours after tail fin injury. Left, bright field; middle, fluorescence; right, merge. Yellow rectangle indicates the analyzed tail region. (F-H) Quantification of neutrophil migration of WT siblings, WT siblings injected with human G226E mRNA, srp54+/−, and srp54+/− zebrafish injected with human G226E mRNA. (F) Total number of mpo+ cells, and (G) mpo+ cells at wound. (H) Fraction of cells migrating toward the injury site. Images were acquired with a Point Scanning Confocal Leica SP5-II-MATRIX microscope (original magnification, ×10). Neutrophils were automatically counted using ImageJ software (we used 3 biological replicates with at least 2 to 3 larvae per replicate). Student t test was used for statistical analysis. ns, not significant. *P < .05; **P < .01; ***P < .005. Bar plots and horizontal lines in the graph represent the mean value of the replicates. Error bars indicate the standard deviation of the mean.

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