Alexa fluor 700 anti mouse cd45 antibody

Zebrafish and mouse embryonic quantitative RT-PCR (qPCR) were conducted according to a previous method^{49 (link)}. Zebrafish embryonic total RNA isolation (Roche, 11667165001) and cDNA synthesis (Thermo Fisher Scientific, 18080051) were performed as previous described^{49 (link)}. HuC-GFP⁺ and coro1a-DsRed⁺ cells were sorted from 3 dpf zebrafish midbrains by flow cytometry. Mouse mesencephalon cell suspensions were incubated with FITC anti-mouse/human CD11b antibody (Elabscience, E-AB-F1081C, 5:100 in blocking buffer containing 5% FBS in dPBS) and Alexa Fluor 700 anti-mouse CD45 antibody (BioLegend, 103128, 2:100 in blocking buffer containing 5% FBS in dPBS) at 4 °C for 1 h. Then, CD11b^hiCD45^lo cells were sorted by flow cytometry. These harvested cells and primary microglial cells were applied to generate cDNA libraries by using REPLI-g WTA Single Cell Kit (QIAGEN, 150063). Relative mRNA levels were determined using the Eppendorf qPCR machine and gapdh or ef1α was used as the control reference. Samples were run in duplicate or triplicate, and the data are presented as mean ± SEM. Genes primers for qPCR assay were listed in Supplementary Table 1.