Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rb1 and compound K improve insulin signaling and inhibit ER stress-associated NLRP3 inflammasome activation in adipose tissue

doi: 10.1016/j.jgr.2015.11.002

Figure Lengend Snippet: Rb1 and CK modulate IRS-1 phosphorylation in the presence of high glucose concentrations. Epididymal adipose tissue was separated after the mice were scarified. The tissue was pretreated with Rb1, CK, or TUDCA at given concentrations, followed by stimulation with high glucose (33mM) for 24 h with or without insulin treatment for an additional 30 min. (A,B) Serine phosphorylation of IRS-1 (S307) and tyrosine phosphorylation IRS-1 (PY99), respectively, were determined by western blot. (C) The level of PI3K in the supernatant of lysed tissue was assayed with an ELISA kit. All results were derived from three independent experiments for the western blot or expressed as the mean ± SD ( n = 4) for ELISA. * p < 0.05 versus high glucose-only treatment; ** p < 0.05 versus the indicated treatment. CK, ginsenoside compound K; ELISA, enzyme-linked immunosorbent assay; SD, standard deviation; TUDCA, tauroursodeoxycholic acid.

Article Snippet: Monoclonal antibodies were procured from the cited commercial sources: anti-TXNIP (NBP1-54578) and anti-NLRP3 (NBP2-12446), Novus Biologicals (Littleton, CO, USA); anti-PERK (#3192), Cell Signaling Technology (Beverly, MA, USA); anti-phospho-PERK (Thr 981; sc-32577) and anti-phospho-IRS-1 (PY99; sc-7020), Santa Cruz Biotechnology, Inc. (Dallas, TX, USA); anti-phospho-IRE1α (S724; ab104157) and anti-IRE1α (ab37073), Abcam (Cambridge, MA, USA); anti-phospho-IRS-1 (Ser307; BS4104), anti-IRS-1 (BS1408), anti-phospho-Akt (T308; BS40080), anti-Akt (BS1810), anti-GAPDH (AP0063), and goat anti-mouse IgG (H+L) horseradish peroxidase (HRP; BS12478), Bioworld Technology (St. Paul, MN,USA).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Derivative Assay, Standard Deviation