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Figure 3: Inhibition of the activity of gamma-secretase after transfection with a dominant negative form of presenilin 1, presenilin 1D385A. (a) The expression of presenilin 1 D385A was detected in the stable transfected cell lines by Western blot. The presenilin 1 D385A protein accumulated as a ~42 kDa fragment (full length) since the mutation-reduced gamma-secretase processing of itself. (b) The activities of gamma-cleavage in G10 and E12 cells were measured using a luciferase-based gamma-secretase reporter system. Data were presented as percent activity where 100% activity corresponds to the reporter gene activity obtained in the nontransfected BEL7404 cells. (c) Cell viabilities of G10, E12, and nontransfected BEL7404 cells were assayed for five consecutive days using the cell counting kit 8 methods. The results represent two independent experiments in triplicate. (d) G10, E12, and nontransfected BEL7404 cells were plated per well in six-well plates for soft agar colony assay, and experiments were performed in duplicates. Data are presented as percent value where 100% value corresponds to the colony obtained in the vehicle control

Figure 3: Inhibition of the activity of gamma-secretase after transfection with a dominant negative form of presenilin 1, presenilin 1D385A. (a) The expression of presenilin 1 D385A was detected in the stable transfected cell lines by Western blot. The presenilin 1 D385A protein accumulated as a ~42 kDa fragment (full length) since the mutation-reduced gamma-secretase processing of itself. (b) The activities of gamma-cleavage in G10 and E12 cells were measured using a luciferase-based gamma-secretase reporter system. Data were presented as percent activity where 100% activity corresponds to the reporter gene activity obtained in the nontransfected BEL7404 cells. (c) Cell viabilities of G10, E12, and nontransfected BEL7404 cells were assayed for five consecutive days using the cell counting kit 8 methods. The results represent two independent experiments in triplicate. (d) G10, E12, and nontransfected BEL7404 cells were plated per well in six-well plates for soft agar colony assay, and experiments were performed in duplicates. Data are presented as percent value where 100% value corresponds to the colony obtained in the vehicle control