Journal: JCI Insight
Article Title: Induction of antiinflammatory purinergic signaling in activated human iNKT cells
Figure Lengend Snippet: Kinetics of changes in the transcription of purinergic genes, cytokines, and transcription factors following activation of cultured human iNKT cells. Cells were collected in RLT lysis buffer (Qiagen) at various times after activation by TCR engagement (5 μg/ml plate-bound αCD3 Ab) and transcripts were quantified by RT-PCR. The results are representative of duplicate experiments. Gene names and corresponding proteins are: ADORA2A , adenosine A2A receptor; CD38 , cyclic ADP-ribose hydrolase 1/NAD + nucleosidase; CD39 , ectonucleoside triphosphate diphosphohydrolase 1; ENPP1 , ectonucleotide pyrophosphatase/phosphodiesterase 1; CD73 , ecto-5-nucleotidase; P2RX7 , P2X7 receptor; PANX1 , pannexin 1; ENT1 , equilibrative nucleoside transporter 1; ADA ( ADA1 ), adenosine deaminase; CD69 , early lymphocyte activation antigen; IFNG , interferon γ; TBX21 ( TBET ), T-box 21; GATA3 , GATA binding protein 3.
Article Snippet: Cultured human iNKT cells were harvested, lysed with RLT Plus lysis buffer (Biolegend), and RNA was extracted using Qiagen Allprep DNA/RNA Micro columns as described by the manufacturer. cDNA was synthesized from RNA samples using Qiagen QuantiTect Reverse Transcription Kits according to the manufacturer’s protocol.
Techniques: Activation Assay, Cell Culture, Lysis, Reverse Transcription Polymerase Chain Reaction, Binding Assay