Pinocembrin inhibited receptor for advanced glycation end products (RAGE)-induced p38 mitogen-activated protein kinase (MAPK)-MAPKAP kinase-2 (MK2)-heat shock protein 27 (HSP27) pathway of human neuroblastoma SH-SY5Y cells overexpressing the Swedish mutant form of human APP (APPsw) in the presence of copper. (A) Images of p38MAPK-MK2-HSP27 pathways performed by high content analysis on the ArrayScan high-content screening (HCS) Reader using the Cytoplasm to Nucleus Translocation BioApplication. APPsw cells were treated plus copper as described in the Methods, prior to fixation, permeabilization, Hoechst nuclear staining, and immunolabeling with antibodies targeting phospho-p38MAPK, phospho-MK2, and phospho-HSP27. Each column reflects images collected from the respective fluorescent channels. (B,C) Values of Mean_CircRingAvgIntenDiff describing the capacity translocation of cytosolic phospho-p38 to the nucleus and nuclear phospho-MK2 to the cytoplasm. (D) Cytosolic Mean_AvgInten value illustrating the expression of phospho-HSP27. Data are expressed as mean ± SEM, n = 6, ***P < 0.001 vs control, #P < 0.05, ##P < 0.01, ###P < 0.001 vs copper.
Liu et al. BMC Medicine 2012 10:105 doi:10.1186/1741-7015-10-105