Humanized transgenic zebrafish express human CXCL12, KITLG, and CSF2. Transgenic zebrafish expressing human cytokines were generated by co-injecting tol2 mRNA and plasmids (A and C). (A) A cartoon of the construct used to make the transgenic zebrafish expressing human CXCL12 (hCXCL12) along with tagBFP under the zebrafish cxcl12 promoter. (B) Representative image of a transgenic zebrafish expressing human CXCL12 in the posterior hemal arc near the tip of the tail at 3 days post-fertilization (dpf). CXCL12 expression continues to progress anteriorly through the hemal arc (representative image shows 8 dpf larvae). (C) A cartoon of the constructs used to make the tet-inducible human SCF/KITLG and GM-CSF/CSF2 expressing zebrafish. (D) Representative image of the human SCF/KITLG (hKITLG) and GM-CSF/CSF2 (hCSF2)-expressing zebrafish. Image specification: magnification = 5x, numerical aperture = 0.16. (E) Representative western blot showing expression of human GM-CSF/CSF2 (hCSF2) and SCF/KITLG (hKITLG) in transgenic zebrafish. S1 and S2 denote samples from the transgenic larvae, and C1 and C2 are samples from control casper larvae. SCF/KITLG: stem cell factor/KIT ligand; GM-CSF/CSF2: granulocyte-monocyte colony- stimulating factor/colony-stimulating factor 2.
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