Journal: Chinese Medical Journal

Article Title: Role of the Ca 2+ -Calcineurin-Nuclear Factor of Activated T cell Pathway in Mitofusin-2-Mediated Immune Function of Jurkat Cells

doi: 10.4103/0366-6999.223855

Figure Lengend Snippet: Effect of MFN2 on intracellular calcium, calcineurin expression, and NFAT activity in Jurkat cells. Jurkat cells were transfected with LV-MFN2 and MFN2-RNAi and then stimulated with PMA (50 ng/ml) plus ionomycin (1 mmmol/L) for various lengths of time. (a and c) Intracellular calcium was measured by FACS with the fluorescent probe Fluo-3/AM. (b) Calcineurin activity was measured using a calcineurin assay kit. (d) NFAT activity in Jurkat cells was measured by ELISA. * P < 0.05, † P < 0.01 versus the control group; ‡ P < 0.05, § P < 0.01 versus the control-RNAi or LV-GFP group. LV: Lentiviral vector; MFN2: Mitofusin-2; GFP: Green fluorescent protein; NFAT: Nuclear factor of activated T cells; PMA: Phorbol myristate acetate.

Article Snippet: Nuclear extract and TransAM NFAT kits were obtained from Active Motif (Carlsbad, CA, USA).

Techniques: Expressing, Activity Assay, Transfection, Enzyme-linked Immunosorbent Assay, Control, Plasmid Preparation

Journal: Chinese Medical Journal

Article Title: Role of the Ca 2+ -Calcineurin-Nuclear Factor of Activated T cell Pathway in Mitofusin-2-Mediated Immune Function of Jurkat Cells

doi: 10.4103/0366-6999.223855

Figure Lengend Snippet: Regulation of calcineurin expression reverses MFN2-induced NFAT activation in Jurkat cells. After various pretreatments, Jurkat cells were stimulated with PMA plus ionomycin for 24 h. Jurkat cells were pretreated with MFN2-RNAi and then LV-calcineurin, and calcineurin activity was measured by commercial assay kit (a) and NFAT activity was determined by ELISA (c). Jurkat cells were pretreated with LV-MFN2 and then FK-506, and calcineurin activity (b) and NFAT activity (d) were measured. * P < 0.05 vs. the control group; † P < 0.05 vs. the control-RNAi or LV-GFP group; ‡ P < 0.05 vs. the LV-MFN2 or MFN2-RNAi group. LV: Lentiviral vector; MFN2: Mitofusin-2; GFP: Green fluorescent protein; NFAT: Nuclear factor of activated T-cell. P/I: PMA (phorbol myristate acetate)/ionomycin; ELISA: Enzyme-linked immunosorbent assay.

Article Snippet: Nuclear extract and TransAM NFAT kits were obtained from Active Motif (Carlsbad, CA, USA).

Techniques: Expressing, Activation Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Control, Plasmid Preparation

Journal: Free radical biology & medicine

Article Title: The HIV-Tat protein interacts with Sp3 transcription factor and inhibits its binding to a distal site of the sod2 promoter in human pulmonary artery endothelial cells

doi: 10.1016/j.freeradbiomed.2019.12.015

Figure Lengend Snippet: Using an Sp-specific oligonucleotide-binding ELISA (TransAm, Active Motif), we measured the levels of active Sp1 (Panel A) and Sp3 (Panel B) in the nucleus of mock transfected (Tat(−)) or pCP2-Tat101 transfected (Tat(+)) HPAEC. The signal was normalized to μg of nuclear protein. Mean ± SEM of biological replicates, n = 3, (***p ≤ 0.001 by unpaired student’s T-test).

Article Snippet: DNA-binding ELISAs for active Sp1 and Sp3 were carried out on nuclear extracts prepared as previously described [ 21 ] using Sp1 and Sp3-specific TransAM kits (Active Motif) according to manufacturer’s protocol.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Transfection

Journal: Molecular Vision

Article Title: Cis-urocanic acid inhibits SAPK/JNK signaling pathway in UV-B exposed human corneal epithelial cells in vitro

doi:

Figure Lengend Snippet: DNA binding of c-Fos and c-Jun subunits of the transcription factor AP-1 heterodimer. Binding of c-Fos ( A ) and c-Jun ( B ) to DNA. Results are presented as mean optical density (OD) ± SEM cis-UCA concentration was 100 µg/ml. Seven parallel samples were measured in control and cis-UCA, and nine parallel samples in UV and cis-UCA + UV treatments. *p<0.05; **p<0.01 (Mann–Whitney).

Article Snippet: To detect the binding of AP-1 and NF-κB to DNA, c-Fos and c-Jun TransAM TM kits (Active Motif, Rixensart, Belgium), and NF-κB p65 ELISA Kit (Enzo Life Sciences, Farmingdale, NY) were used.

Techniques: Binding Assay, Concentration Assay, Control, MANN-WHITNEY