Mouse monoclonal anti jnk

1×10 6 uninfected or KSHV-infected cells were lysed, subjected to electrophoresis and transferred to nitrocellulose membranes, as previously described (Gilardini Montani et al., 2018a) . Membranes were blocked in PBS-0.1% Tween 20 solution containing 3% BSA, probed with specific antibodies and developed using ECL Blotting Substrate (Advansta). The following antibodies were used: rabbit polyclonal anti-PARP1 (1:500; Cell Signaling Technology, 9542), mouse monoclonal anti-catalase (1:100; Santa Cruz Biotechnology, sc-271803), rabbit polyclonal anti-p-JNK (1:300; Cell Signaling, 4668), mouse monoclonal anti-JNK (1:100; Santa Cruz Biotechnology sc-7345), mouse monoclonal anti-p-Bcl2 (1:100; Santa Cruz Biotechnology sc-293128), mouse monoclonal anti-Bcl2 (1:100; Santa Cruz Biotechnology sc-7382), rabbit polyclonal anti-LC3B (1:1000; Novus Biologicals, NB100-2220SS), mouse monoclonal anti-SQSTM1 (1:500; BD Transduction Laboratories, 610883), rabbit polyclonal anti-ATG5 (1:500; Cell Signaling Technology, 2630), and anti-beta actin (1:10000; Sigma Aldrich, A5441). Goat anti-mouse IgG-HRP and anti-rabbit IgG-HRP (1:10.000 Santa Cruz Biotechnology Inc) were used as secondary antibodies.

The study protocol was approved by the Animal Care and Use Committee of Kunming Medical University. Male 4-week-old Sprague Dawley rats (N = 80, 200 ± 20 g) were purchased from the Animal Center of Kunming Medical University. The rats were housed at 23° ± 2°C and 55 ± 2% humidity. The animals had free access to tap water and normal rat diet prior to the start of the experiment. Ginsenoside Rg1 of >98% purity measured via high-performance liquid chromatography was obtained from the College of Pharmacy of Kunming Medical University. STZ was obtained from Sigma Chemical Co. (St. Louis, MO, USA). The bicinchoninic acid (BCA) protein assay kit was obtained from TIANGEN Biotech (Beijing) Co., Ltd. Glucose and insulin enzyme-linked immunosorbent assay (ELISA) kits were purchased from Bio-Swamp Life Science Lab, China. Mouse monoclonal anti-JNK, anti-phospho-JNK (p-JNK), anti-caspase-3, anti-BCL-2-associated X protein (BAX), anti-B cell leukemia/lymphoma 2 (BCL-2), and anti-b-actin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Goat anti-mouse antibody labeled with horseradish peroxidase was obtained from Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China.