MSCs were starved for 15-18 hours in serum-free DMEM/LG containing 0.5% fetal bovine serum (Thermo, Logan, UT), then treated with 2 μg/ml IL-1β inhibitor IL-1RA (PeproTech, NJ, USA) for 2 hours prior to cytokine stimulation. The LFA-1/ICAM-1 inhibitor lovastatin (Cayman Chemical, USA) was added to the cell coculture at a concentration of 50 μM. MAPK p38 inhibitor SB 203580 (5 μM) (Tocris, UK) was added to the cell culture after IL-1β stimulation. According to our previous study, MSCs treated with 100 ng/ml human recombinant IL-1β for 18 hours significantly enhanced migration without affecting cell viability and cell proliferation [10 (link), 32 (link)]. At the indicated time, cells were incubated for 30 minutes with 100 ng/ml human recombinant IL-1β (PeproTech, NJ, USA) in the continued presence of these inhibitors.
HUVECs were starved for 3 hours in serum-free DMEM/F12 containing 1% bovine serum albumin (Sigma, MO, USA), then treated with 100 ng/ml IL-1β for 6 hours.
HUVECs were starved for 3 hours in serum-free DMEM/F12 containing 1% bovine serum albumin (Sigma, MO, USA), then treated with 100 ng/ml IL-1β for 6 hours.
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