rig i reporter cell line  (InvivoGen)

Journal: ACS Omega

Article Title: Morutamins A–L from Morus alba L. Root Bark with Inhibitory Effects on Epidermal Growth Factor Receptor and Toll-Like Receptor-Mediated NF-κB/AP‑1 Activation

doi: 10.1021/acsomega.5c11810

Figure Lengend Snippet: Inhibitory effects of isolated compounds on TLR agonist-induced NF-κB/AP-1 activation in HEK-Blue reporter cells. New compounds are shown in green. HEK-Blue cells expressing human (A) TLR2/1, (B) TLR3, (C) TLR4, (D) TLR5, (E) TLR7, or (F) TLR8 were pretreated with MAE (50 μg/mL) or test compounds (50 μM) for 1 h, followed by stimulation with specific agonists: Pam 3 CSK 4 (10 ng/mL), poly­(I:C) (1 μg/mL), LPS (1 μg/mL), FLA-ST (10 ng/mL), imiquimod (1 μg/mL), or R848 (1 μg/mL). After 24 h, SEAP activity, indicating NF-κB/AP-1 pathway activation, was measured at 620 nm. Data are presented as mean ± SEM ( n = 3) from three independent experiments. Significant differences were determined at * p < 0.05, ** p < 0.01, *** p < 0.001 versus vehicle group.

Article Snippet: HEK-Blue hTLR reporter cell lines (InvivoGen) were cultured in 100 mm cell culture dishes with Dulbecco’s Modified Eagle’s Medium (DMEM; HyClone) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Invitrogen), 100 U/mL penicillin-streptomycin (Gibco), and 0.2% Normocin (InvivoGen).

Techniques: Isolation, Activation Assay, Expressing, Activity Assay