ON and OFF SACs identified by scRNA-seq

The goal of this experiment was to investigate the heterogeneity of SACs via scRNA-seq P0 ChATCre:Thy1-STP-YFP (line#15) retinas were digested with papain (Worthington, LS003126) at 37°C for 10 mins. Retinas were then dissociated and triturated into single cell suspensions. YFP labeled SACs were further isolated by FACS. Single SAC suspensions were loaded into 10X Chromium Single Cell A Chips at a concentration of ~1,000 cells/µL. Single cell libraries were prepared using the Chromium 3' v2 platform (10X Genomics, Pleasanton, CA) as previously described (Peng et al., 2019). Libraries were sequenced on the Illumina HiSeq 2500 (Paired-end reads: Read 1, 26bp, Read 2, 98bp).本实验旨在通过单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）探究SACs的细胞异质性。取出生后第0天（P0）的ChATCre:Thy1-STP-YFP（品系#15）小鼠视网膜，使用木瓜蛋白酶（Worthington，货号LS003126）在37℃下消化10分钟；随后将视网膜解离并吹打成单细胞悬液，通过荧光激活细胞分选（Fluorescence Activated Cell Sorting, FACS）进一步分离YFP标记的SACs。将单细胞悬液以约1000个细胞/微升的浓度上样至10X Chromium单细胞A芯片，参照已发表方法（Peng等，2019），采用Chromium 3' v2测序平台（10X Genomics公司，加利福尼亚州普莱森顿）构建单细胞文库，最终在Illumina HiSeq 2500测序仪上进行双端测序（Read 1为26bp，Read 2为98bp）。