The active component(s) in ACM is heat labile and smaller than 3 kDa.
A, Primary microglia were treated with ACM or
heated ACM (70°C for 15 min) for 12 h. B, Primary
microglia were treated with 20 ng/ml IFN-γ for 12 h in the absence or presence
of ACM or heated ACM. C, BV2 cells were treated with
ACM components smaller than 3 kDa (3↓) or ACM components larger than 3 kDa
(3↑) for 12 h. D, BV2 cells were treated with
20 ng/ml IFN-γ for 12 h in the absence or presence of ACM components smaller
than 3 kDa (3↓) or ACM components larger than 3 kDa (3↑). ACM
components were fractionated as described in Materials and Methods. HO-1
(A, C) and iNOS
(B, D) expression
were measured by Western blotting. The data are representative of three independent
experiments.