Figure 2

The Role of Pez in Macrophage Wound Recruitment Is Cell Autonomous and Dependent upon the FERM Domain

(A) Macrophage-specific expression of Pez-RNAi (TRiP constructs) impairs inflammatory recruitment to wounds (images 1 h post-wounding). Scale bars represent 10 μm. Wound margin is denoted by dashed red line.

(B) Pez-RNAi significantly reduces macrophage recruitment to wounds compared to control (n ≥ 21 wounded embryos/genotype; Kruskal-Wallis with Dunn’s multiple comparisons).

(C) Pez-sfGFP expression in macrophages (outlined in red) co-expressing either control RNAi or either Pez-RNAi. Scale bars represent 10 μm.

(D) Both RNAi lines significantly reduce macrophage Pez-sfGFP intensity levels (n = 18 cells from 6 embryos/genotype; Kruskal-Wallis with Dunn’s multiple comparisons).

(E) UAS-Pez expression constructs. FERM domain and PTP domains noted and deletions depicted. For phosphatase dead construct (UAS-Pez^ΔPD), the mutated cysteine is noted. Adapted from Poernbacher et al.¹⁷

(F) Images of wounded Pez² embryos with macrophage-specific expression of indicated Pez constructs, 1 h post-ablation. Scale bar represents 20 μm. Wound margin is marked by dashed red line.

(G) Macrophage-specific expression of UAS-Pez, UAS-Pez^ΔPD, and UAS-Pez^ΔPTP (but not Pez^ΔFERM) is sufficient to rescue Pez² wound recruitment defect (n ≥ 13 wounded embryos/genotype; one-way ANOVA with Dunnett’s multiple comparisons to Pez²).

(H) Quantification of meandering index reveals specific expression of Pez rescues the inflammatory chemotaxis of Pez² macrophages (n ≥ 42 cells from n ≥ 5 wounded embryos/genotype; unpaired t test).

All error bars are mean ± SD. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.005, and ^∗∗∗∗p < 0.001. See also Figure S2 andVideo S4.