RNA-seq analysis of neonatal mouse cochlear supporting cells [NonTreated_JM]

This study examined transcripts that are enriched in neonatal mouse cochlear supporting cells at postnatal day 1 and postnatal day 6. Supporting cells were purified by FACS sorting for GFP fluorescence from the cochleas of transgenic mice in which a BAC including the LFng locus drives the expression of GFP. Two replicates of GFP+ supporting cells were compared with all other cochlear cell types that were GFP-. We performed this experiment at two different ages, postnatal day 1 and postnatal day 6. mRNA profiles of supporting cells (GFP+) and all other cochlear cell types (GFP-), two replicates each, at P1 and P6 mice were generated by deep sequencing using Illumna TruSeq.本研究分析了出生后第1天（P1）与出生后第6天（P6）的新生小鼠耳蜗支持细胞中富集的转录本。研究人员从携带可驱动绿色荧光蛋白（Green Fluorescent Protein, GFP）表达、且包含LFng基因座的细菌人工染色体（Bacterial Artificial Chromosome, BAC）的转基因小鼠耳蜗中，通过荧光激活细胞分选（Fluorescence-Activated Cell Sorting, FACS）基于GFP荧光信号纯化获得支持细胞。将两份GFP阳性（GFP+）支持细胞的生物学重复样本，与所有其他GFP阴性（GFP-）的耳蜗细胞类型进行对照比较。本研究分别在出生后第1天和第6天两个不同年龄阶段开展此项实验，对P1和P6龄小鼠的支持细胞（GFP+）及其他所有耳蜗细胞类型（GFP-）各设置两份生物学重复，采用Illumina TruSeq建库试剂盒进行深度测序，以获取其mRNA表达谱。