The mRNAs alteration induced by Neat1 knockdown in HT22 cells

We knocked down the lncRNA-Neat1 by RNA interference in HT22 cells, which were immortalized from the mice hippocampal neurons. Then we measured the mRNA alteration induced by Neat1 knockdown through the next generation sequencing technique (RNA-seq). A total of 18,309 transcripts were detected in the current sequencing study, in which 593 transcripts were selected out according to the criteria: |log2(fold change)| > 1, P < 0.05 (22 up-regulated and 571 down-regulated). To explore whether the mRNA change induced by Neat1 knockdown in a stressed culture condition was different from that in normal condition, the cells were dealt with OGD and followed by RNA-seq. A total of 18,502 transcripts were detected, in which 121 transcripts were significantly up-regulated and 126 transcripts were significantly down-regulated according to the aforementioned filter rules. Overall design: We carried out RNA-seq to detect the mRNA change induced by Neat1 knockdown in normal culture condition and stressed condition.

本研究以小鼠海马神经元永生化的HT22细胞为模型，通过RNA干扰（RNA interference, RNAi）敲低长链非编码RNA（long non-coding RNA, lncRNA）-Neat1。随后利用下一代测序技术（next generation sequencing, RNA-seq）检测Neat1敲低所诱导的mRNA表达变化。本测序研究共检测到18309条转录本，根据筛选标准|log₂(倍数变化)|>1且P<0.05，共筛选出593条差异表达转录本，其中22条上调、571条下调。为探究Neat1敲低在应激培养条件下诱导的mRNA变化与正常培养条件下是否存在差异，本研究对细胞进行氧糖剥夺（oxygen-glucose deprivation, OGD）处理后开展RNA-seq测序。本批次测序共检测到18502条转录本，依据前述筛选标准，共获得121条上调差异转录本与126条下调差异转录本。实验整体设计：本研究通过RNA-seq分别检测正常培养与应激培养条件下，Neat1敲低所诱导的mRNA表达变化。