ZFIN Image: Candel et al., 2014, Fig. 5

Image

Figure Caption

Fig. 5

Tnfa and Tnfr2 deficiencies result in the Duox1-derived H₂O₂ production by keratinocytes.

Zebrafish one-cell krt18:RFP (A), wild-type (B, C), or mpx:eGFP (D–G) embryos were injected with standard control (Std), Tnfr2, Tnfa, Duox1/p53, and/or Lyn MOs. (A) The expression of the duox1 gene was measured by RT-qPCR in FACS-sorted keratinocytes from 72 hpf wild-type and Tnfr2-deficient larvae. (B, C) Wild-type and Tnfr2-deficient larvae were dechorionated at 24 hpf and treated by immersion in 100 μM DPI or vehicle alone (DMSO) for 24 h and then labeled with 50 μM acetyl-pentafluorobenzene sulphonyl fluorescein. Representative images of green channels of Std and Tnfr2 morphants are shown. Note that single keratinocytes are labeled with the H₂O₂ probe in Tnfr2-deficient larvae (inset). (D–G) Rescues with Duox1 (D, E) and Lyn (F, G) MOs at 72 hpf. The differences in the neutrophil distribution (D, F) and quantification of neutrophil mobilization from the CHT to the skin in the indicated number of larvae per group from three different experiments (E, G) are shown. The mean ± S.E.M. for each group is shown. Scale bars, 100 μm. ns, not significant. ***p<0.001.

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ PLoS Biol.