Anti tim 3 mab clone f38 2e2

HUVECs were cultured overnight in complete medium, and then incubated in serum-free medium for 12 hours, followed by stimulation with a range of ox-LDL concentrations (0, 1, 50, and 100 µg/mL) for 48 hours. HUVECs were collected for flow cytometric analysis. Selected cell cultures were treated with 1,000 ng/mL recombinant Tim-3 (2365-TM-050, R&D Systems, Minneapolis, MN, USA), 10 mg/mL anti-Tim-3 mAb (clone F38-2E2, Biolegend, San Diego, CA, USA), NF-κB inhibitor BAY 11-7082 (30 µM), ERK1/2 inhibitor U0126 (30 µM), JNK signaling pathway inhibitor SP600125 (30 µM), P38 signaling pathway inhibitor SB202190 (30 µM), or PI3K signaling pathway inhibitor LY294002 (30 µM).

Freshly isolated DSCs were cultured overnight in complete medium and further incubated in serum-free medium for 12 h, followed by stimulation with a range of concentrations of the TLR ligands peptidoglycan from Staphylococcus aureus (pGN-SA; 08K03-SV, InvivoGen, San Diego, CA, USA), poly(I:C) (11C21-MM, InvivoGen), and LPS (13I06-MM, InvivoGen) for 48 h. DSCs were collected for flow cytometric analysis. To some wells were added 1000 ng/ml recombinant Tim-3 (2365-TM-050, R&D Systems, Minneapolis, MN, USA), 10 μg/ml anti–Tim-3 mAb (clone F38-2E2, Biolegend, San Diego, CA, USA), JNK signaling pathway inhibitor SP600125 (30 μM), P38 signaling pathway inhibitor SB202190 (30 μM), NF-κB inhibitor BAY 11-7082 (30 μM), or MEK1/2 inhibitor U0126 (30 μM).