After culturing and expanding electroporated cells for 10 days, 0.5–1 million cell pellets were harvested for genomic DNA and RNA extraction. Genomic DNA was extracted using QuickExtract lysis buffer (Qiagen, SS000035-P) following manufacturer’s recommendations. PCRs were performed for each ...
Therefore, we use single cell RNA sequencing (scRNA-seq) to reveal distinct immune microenvironment in primary and brain metastatic lesion of NSCLC.Methods:Eight primary and nine brain metastatic NSCLC tumor samples were obtained through surgical resection. Immune cells (CD45+) were enriched through ...
Cell cycle was analyzed using Cell-Clock cell cycle assay on BECs by the treatment with serum depletion or GCDC for 4 days with or without knockdown ofIFIT3usingsiRNA. G1/S arrest was induced by the treatment with serum depletion or GCDC (p < 0.05) (Fig.2E). G1/S arrest was ...
Human induced pluripotent stem cells (iPSCs) are used to generate models of human diseases that recapitulate the pathogenic process as it occurs in affected cells. Many differentiated cell types can currently be obtained from iPSCs, but no validated prot
Mus dunni endogenous virus (MDEV) infects a wide variety of cell types from many different species. To take advantage of this broad host range, we have constructed packaging cells (PD223) that produce virions bearing the MDEV envelope. PD223 cells are able to package Moloney murine leukemia ...
2.2. RNA extraction, library preparation and sequencing Total RNA was extracted from purified monocytes using the RNeasy Mini-Kit with DNase treatment (QIAGEN, Germany). The RNA quality and quantity were measured using NanoDrop 2000 (Thermo Fisher Scientific Inc., USA) and Qubit 2.0 RNA Broad Ra...
T Cell-Specific siRNA Delivery Suppresses HIV-1 Infection in Humanized Mice 2008, Cell Citation Excerpt : It has been suggested that for a chronic infection like AIDS, a sustained antiviral state is best achieved by a gene therapy approach where vector-mediated delivery of shRNA to hematopoietic...
RNA sequencing Total RNA was isolated from three independent confluent VDOP cell lines using RNeasy mini kit (Qiagen, Valencia, CA) following the manufacturer’s protocol. An RNA library was prepared for each cell line using the TruSeq RNA sample Prep kit version 2 (Illumina, San Diego, CA,...
RNA extraction was performed using QIAGEN RNEasy kit following manufacturer’s protocol. RNA was eluted in 50 μl of nuclease free H2O and required further concentration for cDNA synthesis. RNA concentration was performed using ZYMO RNA Clean and ConcentratorTM−5 kit. ...
The supernatant obtained in the second round of extraction was combined with that from the first round. The solution containing the cell metabolites was then dried in a vacuum concentrator. The dry residue was reconstituted with 500 uL of methanol and water (80:20, v/v) for subsequent ...