Fig. 1: Apolipoproteins E suppresses CLSP activity.
From: Restoration of the reduced CLSP activity alleviates memory impairment in Alzheimer disease
a ApoE3, E4, adiponectin (ADN), and annexin 2, C-terminally tagged with HA, were overexpressed in F11 neurohybrid cells. Resulting cell lysates were mixed with bacterially produced GST-MycHis- or CLSP-MycHis-conjugated sepharose 4B and incubated at 4 °C overnight. The pulled-down precipitates were immunoblotted using HA and myc antibodies. For reference, inputs including the sepharose 4B beads conjugating GST-MycHis (GST-MH) and CLSP-MycHis (CLSP-MH) and the cell lysates were similarly immunoblotted. b SH-SY5Y cells, transfected with the empty vector or pcDNA3.1/MycHis-V642I-APP, were cultured in the presence of indicated concentrations of CLSP-MycHis. At 48 h, cells were harvested for trypan blue exclusion cell mortality assays (Trypan Blue Test) and cell viability assays using the WST-8 assay kit or staining with calcein AM. Cell lysates were immunoblotted using the APP antibody. c, d SH-SY5Y cells, transfected with the vector or pcDNA3.1/MycHis-V642I-APP, were cultured in media containing 1 nM of GST-MycHis or CLSP-MycHis with/without indicated concentrations of BSA, ApoE3 (c), or ApoE4 (d). At 48 h, cells were harvested for trypan blue exclusion assays. Cell lysates were immunoblotted using the APP antibody.
