C100 and Aβ protein production in vitro after plasmid transfection.

Analysis of C100 and Aβ protein production in vitro after transfection with plasmids expressing Aβ40-GFP, Aβ42-GFP, C100-GFP or C100^V717F-GFP in cell homogenates (A) and media (B). C100 protein was only detected in cell homogenates after transfection with C100-GFP and C100^V717F-GFP plasmids. No Aβ was detected in cell homogenates after transfection with any plasmid. APP^SWE brain homogenate was used as a positive control for Aβ detection. Strong GFP protein expression indicated successful transfection using all plasmids and β-actin was used as a loading control. Immunoprecipitation of Aβ from the cell culture media revealed Aβ in the media from cells transfected with C100-GFP and C100^V717F-GFP plasmids. Control refers to non-transfected cells.