Hek blue tlr7

Human embryonic kidney (HEK) 293 cells stably expressing human TLR7 or TLR9 cell lines, HEK-Blue TLR7, HEK-Blue TLR9, and its parental cell line HEK-blue Null1 cells were obtained from Invivogen (San Diego, CA, United States) and used to access TLR-mediated NF-κB activation by measuring the SEAP activity (Zhang et al., 2018 (link)). Briefly, 3 × 10⁵ cells/mL HEK-Blue TLR7, HEK-Blue TLR9, or HEK-blue Null1 cells were seeded into 96-well plates for 48 h and then treated with 10 μM IMQ or 1 μM CpG685 in the presence or absence of different concentrations of HJ901. Additionally, the cells were treated with HJ901 in the presence or absence of various concentrations of IMQ or CpG685. These cells were also incubated with 10 μM IMQ or 1 μM CpG685 for 0, 2, 4, 6, and 12 h, and subsequently treated with or without HJ901. At the end of treatment, 20 μL of the culture supernatant was removed from each treatment and tested for SEAP activity using 180 μL of Quanti-Blue substrate following the manufacturer’s protocol (Invivogen). TLR7 or TLR9-mediated NF-κB activation can be assessed by measuring SEAP activity.

Unmodified 4T1 mammary carcinoma cells were obtained from Dr. F.R. Miller (WSU, School of Medicine, Detroit, MI) and were cultured as described in [10 (link)]. 4T1-EGFP stable cell line was obtained by a transfection of 4T1 cells with the lentiviral vector pLV-neo-EGFP followed by FACS-sorting.
K562 human erythromyeloid and YAC-1 mouse lymphoma cell lines are from ATCC. HEK-Blue® cells stably transfected with inducible SEAP reporter gene under NF-κB-dependent promoter and human Toll-like receptors 2, 3, 4, 5, 7, 8 or 9, and their co-receptor molecules CD14 and MD2 (HEK-Blue-hTLR2/CD14, HEK-Blue-TLR3, HEK-Blue-TLR4/CD14-MD2, HEK-Blue-TLR5, HEK-Blue-TLR7, HEK-Blue-TLR8, HEK-Blue-TLR9 cells), were obtained from InvivoGen.