Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-7Rα low CD8 + T Cells from Healthy Individuals Are Anergic with Defective Glycolysis.

doi: 10.4049/jimmunol.1901470

Figure Lengend Snippet: FIGURE 3. GATA3 and c-MYC are downregulated in anergic IL-7Ralow EM CD8+ T cells and recovered by IL-2 reversal. To determine how IL-7Ralow

Article Snippet: GATA binding protein 3 knockdown by small interfering RNA Jurkat T cells (American Type Culture Collection, Manassas, VA) were transfected with control (scramble) or GATA binding protein 3 (GATA3) small interfering RNA (siRNA) (59-GCA-GUA-UCA-UGA-AGC-CUAAAC-GCG-A-39; OriGene, Rockville, MD) using a Neon Transfection System (Invitrogen, Carlsbad, CA) in accordance with the manufacturer’s instructions.

Techniques:

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-7Rα low CD8 + T Cells from Healthy Individuals Are Anergic with Defective Glycolysis.

doi: 10.4049/jimmunol.1901470

Figure Lengend Snippet: FIGURE 5. GATA3 knockdown induces T cell dysfunction in terms of cell proliferation, cytokine production, and cell metabolism. Jurkat T cells were transfected with GATA3 siRNA or control siRNA and then harvested after 72 h. (A) GATA3 knockdown was confirmed by immunoblot analysis. (B) Cell growth was monitored by CCK-8. Cells were stimulated with anti-CD3/CD28 Abs for 12 h. Then, CCK-8 solution was added to the cultures, and the cells were then incubated for 24 h. Absorbance at 450 nm was determined by using a microplate reader. (C) Cells were stimulated with anti-CD3/CD28 Abs and then subjected to OCR and ECAR analyses (2 3 105 cells per well). The p values were obtained using the two-tailed Student t test (n = 5). Data were combined from two independent experiments.

Article Snippet: GATA binding protein 3 knockdown by small interfering RNA Jurkat T cells (American Type Culture Collection, Manassas, VA) were transfected with control (scramble) or GATA binding protein 3 (GATA3) small interfering RNA (siRNA) (59-GCA-GUA-UCA-UGA-AGC-CUAAAC-GCG-A-39; OriGene, Rockville, MD) using a Neon Transfection System (Invitrogen, Carlsbad, CA) in accordance with the manufacturer’s instructions.

Techniques: Knockdown, Transfection, Control, Western Blot, CCK-8 Assay, Incubation, Two Tailed Test