Figure 5. Amoeboid mpeg1+ cells in the zebrafish skin are of non-hematopoietic origin and have a metaphocyte transcriptome.

(A) Immunofluorescence on manually dissected scales from adult skin of control and csf1r^DM mpeg1:EGFP +; kdrl-induced-DsRed+ adults (4 mpf). Stars: single-positive (SP) cells; white arrowheads: double-positive (DP) cells. (B) FACS analysis on cells from the adult skin (4 mpf, n = 3 per group) and quantification. GFP⁺DsRed^-=mpeg1+ only, GFP⁺DsRed⁺=mpeg1+/kdrl-induced-DsRed+. (C) PCA analysis showing segregatin based on cell type (PC1) and genotype (PC2). (D) Volcano plot showing gene expression changes between control GFP⁺DsRed⁺ versus GFP⁺DsRed^- cells. Light grey: DGE of all genes, Green: DGE of genes enriched in metaphocytes logFC >2 (Lin et al., 2019); Orange: DGE of genes downregulated in metaphocytes logFC <2 (Lin et al., 2019). (E) Heat map showing the expression of metaphocyte signature genes. (F) Heat map showing the expression of phagocytosis and engulfment genes. (G) Venn diagram showing DGE between the three groups (logFC > |2|; FDR < 0.05).

Figure 5—figure supplement 1. Gating strategy for isolating mpeg1+ cells from juveniles.

(A) FACS sorting strategy showing one representative example for each genotype. (B) Quantification of the length of the fish and the percentage mpeg1+ cells out of live cells ***<0001. Each dot represents one fish.

Figure 5—figure supplement 2. Expression profiles of non-hematopoietic and hematopoietic mpeg1+ cells of control and csf1r^DM juvenile zebrafish.

(A) Graphs showing counts per million (cpm) values for various genes enriched in macrophages. (B) Heat map showing the expression of genes downregulated in metaphocytes compared to neutrophils and LCs (LogFC <2, Lin et al., 2019).