2-ClHDA activates MAPK pathways.

<p>(A) Concentration-dependent activation of the MAPK cascade. BMVEC were incubated with NaOCl or 2-ClHDA or DMSO (used as vehicle for 2-ClHDA delivery) at the indicated concentrations for 3 h. (B) Time-dependent activation of the MAPK cascade in response to 2-ClHDA. BMVEC were incubated with 2-ClHDA (25 µM) for the indicated time periods. After treatment, cells were lysed, aliquots of protein lysates were subjected to SDS-PAGE and transferred to PVDF membranes. Pan- or phospho-specific polyclonal antibodies against p38, JNK1/2, or ERK1/2 were used as primary antibodies. Immunoreactive bands were visualized with peroxidase-conjugated secondary antibodies using the ECL-system. Bar graphs in the right panels show the ratio of optical densities of immunoreactive phosphorylated normalized to non-phosphorylated proteins.