Fig. 1: Construction of zebrafish irg1/acod1 GFP knock-in for tracking the dynamics of macrophage activation in vivo.
a Construction of vector and zebrafish line based on GeneWeld method25,26. b Site-specific insertion of GFP and verification of insertion by a suite of PCR assays. See Supplementary Data 1. c Schematic of the GFP knock-in protein sequence at the targeted locus as verified by Sanger DNA sequencing of F1 founders (#7 and #31) and a F2 progeny. d Dual labeling of activated brain-resident macrophages, also known as microglia, in the larval zebrafish brain after LPS stimulation demonstrates efficacy of the GFP knock-in reporter. Reporter labels LPS-activated microglia with high GFP and conveys heterogenous states as few cells (asterisks) show less activation based on very low GFP. Microglia are labeled by mpeg1:BFP and demarcated by dotted lines. Homeostatic microglia (no LPS) have low baseline GFP expression as expected.
