Next Generation Sequencing to examine common set of transcripts induced in spinal cord neurons by polyPL and mutSOD1-ACM common set of transcripts. Next Generation Sequencing to examine common set of transcripts induced in spinal cord neurons by polyPL and mutSOD1-ACM common set of transcripts

Next Generation Sequencing shows that synthetic polyPL and mutSOD1-ACM alter a large and common set of transcripts in primary ventral spinal cord cultures Overall design: Rat primary spinal cord cultures were incubated for 90 min with long-chain synthetic polyphosphate (polyPL; 100 μM), astrocyte conditioned media (ACM) from mutSOD1G93A astrocytes (hSOD1G93A) or control astrocytes (hSOD1WT). Untreated cells were used as internal controls (control). Total RNA was isolated from cell lysates (in quadruplicate) and mRNA expression was analyzed by strand-specific RNAseq.

下一代测序（Next Generation Sequencing）结果表明，合成聚磷酸盐（polyPL）与突变型SOD1星形胶质细胞条件培养基（mutSOD1-ACM）可在原代腹侧脊髓培养物中改变一大组共有的转录本表达谱。实验设计：将大鼠原代脊髓培养物分别与长链合成聚磷酸盐（polyPL；100 μM）、突变型SOD1G93A星形胶质细胞（hSOD1G93A）的条件培养基（ACM），或野生型人SOD1（hSOD1WT）星形胶质细胞的条件培养基孵育90分钟；以未经过处理的细胞作为内部对照（对照组）。从细胞裂解液中提取总RNA（一式四份），并通过链特异性RNA测序（strand-specific RNAseq）分析mRNA的表达水平。