Subcellular fractionation To determine the cellular localization of protein and mRNA, cytoplasmic and nuclear proteins and RNAs were isolated using PARIS Kit (Thermo) according to the manufacturer’s instructions. The extracted proteins and RNAs were subjected to western blot and RT-qPCR analysis, respe...
Subcellular fraction isolation The nuclear and cytoplasmic fractions were extracted using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific) according to the manufacturer’s instructions. Total RNA from whole-cell lysates or the nuclear and cytoplasmic fractions were isolated using TRIzol...
Subcellular fractionation Nuclear and cytoplasmic separation was performed using the PARIS Kit (Life Technologies, USA) according to the manufacturer’s instructions. Fluorescence in situ hybridization (FISH) The FISH assay was performed in MDA-MB-231 cells according to the specifications of the manufact...
The root cause of PE is believed to be in the placenta, and therefore identification of the factors that affect the development of the placenta has been the focus of recent studies. Researchers have been searching for genes that confer the susceptibilities of this disease. However, most of the...
This was supported by subcellular CCND1 mRNA localization studies utilizing RNA fluorescence in situ hybridization (RNA-FISH) technique. CLSM images showed thatCCND1mRNA was distributed in both nuclei and cytoplasm of HGC cells, explaining the spatial “off–on” switch of fluorescence during GQDs/FA...
5.核质分离:按PARIS™ Kit Protein and RNA Isolation System试剂盒对小鼠肝原代细胞进行核质分离,用实时PCR检测LncRNA-Co7Rik的表达,核内RNA以45S rRNA为内参,细胞质以12S rRNA为内参。6.引物设计:根据生物信息学数据库(Ensembl)查...
Subcellular fractionation assay According to the manufacturer’s instructions, a cytoplasmic and nuclear RNA purification kit (Norgen, Thorold, ON, Canada) was used. Briefly, cells were incubated with a lysis solution to disrupt the cell membrane and centrifuged at 4°C and 1,200 × g to separ...
Jinxin Lin from the College of Biological Science and Technology, Beijing Forestry University, for providing the pBIN2-RFP-HDEL vector in subcellular localization and Dr. Yan Zhu from the Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, for the assistance with ...
Spatiotemporally resolved transcriptomics reveals the subcellular RNA kinetic landscape TEMPOmap combines pulse-chase metabolic labeling with multiplexed three-dimensional in situ sequencing to simultaneously profile the age and subcellular location of individual RNA molecules from thousands of genes to reveal ...
Although the subcellular dynamics of RNA and proteins are key determinants of cell homeostasis, their characterization is still challenging. Here we present an integrative framework to simultaneously interrogate the dynamics of the transcriptome and prot