Figure 3.
Minocycline inhibits p38MAPK-dependent proNGF expression in microglia, BV2 cells.
A
, Cells were plated onto six-well plates and treated with LPS. Note that BV2 cell treated with LPS for 4 h exhibited cellular processes, a characteristic of activated microglial cells.
B
, The level of p-p38MAPK was increased and peaked at 30 min after LPS treatment.
C
, Minocycline (1 and 5 nm) treatment decreased the level of p-p38MAPK at 30 min after LPS treatment when compared with that in cultures treated with LPS only.
D
, Quantitative analysis of Western blots shows that minocycline significantly inhibited p-p38MAPK expression at 30 min after LPS treatment when compared with that in cultures treated with LPS only. Values are mean ± SD of three separate experiments. *p < 0.001.
E
, The level of p-MAPKAPK-2 was increased and peaked at 30 min after LPS treatment.
F
, Minocycline (1 and 5 nm) or SB203580 treatment (1 and 5 μm), an inhibitor of p38MAPK, decreased the level of p-MAPKAPK-2 at 30 min after LPS treatment when compared with that in cultures treated with LPS only.
G
, Quantitative analysis of Western blots shows that minocycline and SB203580 significantly inhibited p-MAPKAPK-2 expression at 30 min after LPS treatment when compared with that in cultures treated with LPS only. Values are mean ± SD of three separate experiments. *p < 0.001.
H
,
I
, NGF mRNA (
H
) and proNGF protein (26 kDa) (
I
) expression were increased and peaked at 2 and 4 h, respectively, after LPS treatment.
J
, Minocycline (1 and 5 nm) and SB203580 (1 and 5 μm) treatment decreased the level of proNGF expression at 4 h after LPS treatment compared with that in cultures treated with LPS only.
K
, Quantitative analysis of Western blots shows that minocycline and SB203580 significantly inhibited proNGF expression at 4 h after LPS treatment when compared with that in cultures treated with LPS only. Values are mean ± SD of three separate experiments. *p < 0.001.