Total RNA was isolated from highly pure, sorted human ILC subsets using the TRIZOL reagent according to the manufacturer’s instructions (Invitrogen). Final preparation of RNA was considered DNA- and protein-free if the ratio of readings at 260/280 nm was ≥1.7. Isolated mRNA was reverse-tr...
After 36 h coculture, TCR-T or IL-21R-TCR-T were stained with TCR-PE antibody and sorted by EasySepTM Release PE Positive Selection Kit (Stemcell Technologies) following the instructions. Single-cell RNA sequencing ScRNA-seq libraries were prepared and sequenced by Genergy Biotechnology (...
The primer extension reactions were performed with a SNaP Shot Dideoxyribonucleoside Triphosphate Primer Extension Kit (Applied Biosystems), according to the manufacturer's instructions. The primer extension reactions were cleaned up by incubation at 37°C for 1 h in the presence of 1 U of SAP, ...
also a plus. · Experience with Power Bi and Microsoft Teams environment preferred. · Superior organizational skills, able to manage and prioritize tasks, meet deadlines, and handle multiple tasks and projects effectively and efficiently. · Exceptional attention to detail, adhere to instructions, ma...
Furthermore, reverse transcription was carried out according to the instructions. Using cDNA as a template and β-actin as an internal reference, the cDNA template obtained by reverse transcription was added to the detection system according to the RT-PCR reagent operating instructions. The ABI7500...
according to the manufacturer’s instructions. Reverse tran- scription and real-time PCR reactions were then carried out using LightCycler-RNA Master Hybridization Probes (one- step procedure) on a LightcyclerR apparatus (Roche Diagnostics, Brussels, Belgium). Primers, standards, and real-time PCR...
Transient transfections were performed according to the manufacturer’s instructions. Western blot Cells were collected by trypsinization and resuspended in NETN150 lysis buffer (150 mM NaCl, 0.5% NP40, 20 mM Tris pH 8.0, 1 mM EDTA) after treated with 0 μM or 5 μM erlotinib ...
and total RNA was isolated according to manufacturer's instructions. For FACS-sorted cell mRNA isolation, RNeasy kit (Qiagen) was used. Complementary DNAs were prepared using SMART MMLV reverse transcriptase (Clontech) and used for PCR with SYBR Green reagents (Qiagen or Quanta Bioscience) on a...
Unconventional protein secretion represents an important process of the inflammatory response. The release of the pro-inflammatory cytokine interleukin (IL)-1β which burst during pyroptosis as a consequence of gasdermin D plasma membrane pore formation,
ST2hi memory-type Th2 cells are identified as a pathogenic subpopulation in eosinophilic airway inflammation. These ST2hi pathogenic Th2 cells produce large amount of IL-5 upon T cell receptor stimulation, but not in response to IL-33 treatment. By contr