Figure 4.

AICAR inhibits LPS-induced mitogen-activated protein kinases (ERK1/2, p38, and JNK1/2) in primary astrocytes: Primary astrocytes were incubated with different concentration of AICAR (0.5 to1 mm) for 2 hr, followed by LPS treatment (1 μg/ml) for 30 min. Cells were washed with chilled PBC and scraped in lysis buffer as discussed in Materials and Methods. Fifty milligrams of total protein was loaded on SDS-PAGE followed by immunoblot analysis with phosphor-specific antibodies against p42/44, JNK1/2, and p38. The same blot was stripped and reprobed with pan antibodies of p42/44, JNK1/2, and p38 for equal loading. Blots are representative of two experiments.