Fig. 2

gpnmb mutants exhibit altered pro- and anti-inflammatory responses after cardiac cryoinjury in zebrafish. A. Illustration of gpnmbbns595 full locus deletion allele generated using the CRISPR/Cas9 technology. RT-qPCR analysis of gpnmb mRNA levels in 5 dpf larvae, dissected caudal fins from adult zebrafish, and 7 dpci injured tissue from adult ventricles, comparing gpnmb+/+ and gpnmb?/?siblings; Ct values listed in Supplementary Table S1; ND= Not detectable. B. Immunostaining for EGFP (macrophages, magenta) with DAPI (DNA marker, blue) counterstaining on sections of Tg(mpeg1:EGFP); gpnmb+/+ and Tg(mpeg1:EGFP); gpnmb?/? cryoinjured ventricles at 7 dpci. C. mpeg1:EGFP+ cell number in gpnmb+/+ and gpnmb?/? BZI tissue (100 ?m) at 7 dpci. D. Immunostaining for Mpx (neutrophils, white) with DAPI (DNA marker, blue) counterstaining on sections of gpnmb+/+ and gpnmb?/? cryoinjured ventricles at 96 hpci. E. Mpx+ cell number in gpnmb+/+ and gpnmb?/? injured tissues at 96 hpci. F. Experimental plan for the bulk RNA-seq analysis performed on the BZI tissue of gpnmb?/? and gpnmb+/+ cryoinjured ventricles at 96 hpci. G. Heatmap showing differentially expressed genes of interest in the BZI tissue of gpnmb?/? and gpnmb+/+ cryoinjured ventricles at 96 hpci. H. GSE analysis plot of TNF-? signaling via NF-kB, the complement system and antigen response from the 96 hpci gpnmb?/? vs gpnmb+/+ transcriptomic analysis. White dotted lines delineate the injured area; white arrowheads point to mpeg1:EGFP+ (B) and Mpx+ (D) cells. Statistical test: Student's t-test (C, E). Scale bars: 100 ?m. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)