Expression profiling of mouse primary prostate cells from WT mice

By single-cell expression profiling and validation, here we report that both basal and luminal-localized cells, particularly the latter, are molecularly heterogeneous. We validate LY6D as a marker linking CR luminal cells to luminal progenitors. Overall design: Primary prostate cells were isolated and luminal prostate cells were FACS-sorted by LY6D+/- SCA1High/Int/low from the lineage (CD31, CD45, Ter119) negative fraction. Total RNA was extracted from two biological replica test. Lexogen Quantseq RNA Sample Prep Kit was used with 10-20 ng of total RNA for the construction of sequencing libraries.本研究通过单细胞表达谱分析与验证，证实基底细胞与腔室定位细胞（尤以腔室细胞为著）均存在分子异质性。我们验证了LY6D可作为连接CR腔室细胞与腔室祖细胞的标志物。 实验整体设计：分离原代前列腺细胞后，从谱系阴性（CD31、CD45、Ter119）组分中，通过LY6D±结合SCA1高/中/低表达水平，对前列腺腔室细胞进行荧光激活细胞分选（Fluorescence Activated Cell Sorting, FACS）。从两份生物学重复样本中提取总RNA，使用Lexogen Quantseq RNA样本制备试剂盒，以10~20 ng总RNA构建测序文库。