Figure 4. p22 phox and Yrk regulate... | Rockefeller University Press

Figure 4.

$Figure 4. p22 phox and Yrk regulate neutrophil reverse migration. (A) Representative images of neutrophil recruitment to wounds in 3-dpf control and cyba morphant (cyba MO) larvae. 1 and 6 hptt, larvae were fixed, and Sudan black staining was used to visualize neutrophils. (B) Quantification of the number of neutrophils at wound in control and cyba morphant (cyba MO) larvae at 1 and 6 hptt. (C) Representative images of neutrophil recruitment to wounds in 3-dpf Tg(mpx:Dendra2) larvae treated with buffer control or yrk MO1 and yrk MO2 at 1 and 6 hptt. Dashed lines border the location of tail transection. (D) Quantification of the number of neutrophils at wounds in C. (E) Schematic of the reverse migration experiment (see Materials and methods). (F) Representative images acquired just before photoconversion, just after photoconversion, and at 4 hpw. 3-dpf Tg(mpx:Dendra2) larvae that were injected with buffer (control) or cyba MO were wounded. (G) Quantification of the number of new neutrophils entering the wounded fin area (bounded by dashed orange squares). (H) Quantification of the fraction of neutrophils remaining in the wounded area. (I) Same experimental setting as in F was used to compare neutrophil reverse migration between control and yrk MO1 larvae. (J) Graph representing the quantification of new neutrophils at the wound 4 hpw. (K) Histogram of the fraction of neutrophils remaining in the fin counted 4 hpw. (n) = number of larvae. Horizontal lines indicate means. Error bars indicate SEM. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001, two-tailed unpaired t test (B, G, H, J, and K) and one-way ANOVA with Dunnett’s post-test (D). Bars, 70 µm.$

p22 phox and Yrk regulate neutrophil reverse migration. (A) Representative images of neutrophil recruitment to wounds in 3-dpf control and cyba morphant (cyba MO) larvae. 1 and 6 hptt, larvae were fixed, and Sudan black staining was used to visualize neutrophils. (B) Quantification of the number of neutrophils at wound in control and cyba morphant (cyba MO) larvae at 1 and 6 hptt. (C) Representative images of neutrophil recruitment to wounds in 3-dpf Tg(mpx:Dendra2) larvae treated with buffer control or yrk MO1 and yrk MO2 at 1 and 6 hptt. Dashed lines border the location of tail transection. (D) Quantification of the number of neutrophils at wounds in C. (E) Schematic of the reverse migration experiment (see Materials and methods). (F) Representative images acquired just before photoconversion, just after photoconversion, and at 4 hpw. 3-dpf Tg(mpx:Dendra2) larvae that were injected with buffer (control) or cyba MO were wounded. (G) Quantification of the number of new neutrophils entering the wounded fin area (bounded by dashed orange squares). (H) Quantification of the fraction of neutrophils remaining in the wounded area. (I) Same experimental setting as in F was used to compare neutrophil reverse migration between control and yrk MO1 larvae. (J) Graph representing the quantification of new neutrophils at the wound 4 hpw. (K) Histogram of the fraction of neutrophils remaining in the fin counted 4 hpw. (n) = number of larvae. Horizontal lines indicate means. Error bars indicate SEM. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001, two-tailed unpaired t test (B, G, H, J, and K) and one-way ANOVA with Dunnett’s post-test (D). Bars, 70 µm.