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Fig. 5.

Effect of compound 4 on LPS-induced MAPK and AP-1 activation. RAW264.7 cells were plated in 100 mm-diameter dishes. After 12 h of seeding, cells were treated with different doses of compound 4 for 1 h, followed by stimulation with 500 ng/ml of LPS for 30 min. Cell lysates (30 µg protein) were prepared and subjected to Western blot analysis by using antibodies specific for (A) IκB-α, phosphorylated forms of ERK1/2, JNK, p38 MAPK and (B) phosphorylated forms of c-Jun. Equivalent loading of cell lysates was determined by reprobing the blots with anti-β-actin, total ERK1/2, JNK or p38 MAPK antibody. (C) Nuclear protein (30 µg protein) was prepared and subjected to Western blot analysis by using antibodies specific for c-Jun and equivalent loading of nuclear protein was determined by reprobing the blots with anti-PARP. The bands were quantified using image analysis software and their relative intensity was expressed as fold against the image of the LPS-stimulated RAW264.7 cells.

Korean J Physiol Pharmacol 2013;17:217-222 https://doi.org/10.4196/kjpp.2013.17.3.217
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