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. 2003 Sep;77(18):9758–9768. doi: 10.1128/JVI.77.18.9758-9768.2003

FIG. 9.

FIG. 9.

FIG. 9.

KLIP1 repressed TK promoter activity in a dose-dependent fashion in a mammalian one-hybrid assay. (A) Luciferase activity in 293 cells either transfected with 1 μg G5Eluc vector alone or cotransfected with 1 μg G5Eluc and 1 μg PM2-KLIP1. GAL4-VP16 was cotransfected with G5Eluc as a positive control. (B) Relative CAT activity in 293 cells cotransfected with 1 μg of GAL4-TK-CAT reporter plasmid and 0, 0.25, 0.5, 075, 1, or 2 μg of PM2-KLIP1. At 2 μg, KLIP1 repressed up to 87% of TK promoter activity. (C) Western blotting assay was carried out with anti-GAL4 DBD antibodies to examine the expression of PM2-KLIP1 fusion protein in 293 cells transfected with different doses of the plasmid DNA cited in (B). α-tubulin expression was used as loading control. (D to F) Relative CAT activity in HeLa (D), BJAB (E) and HUVEC (F) cells cotransfected with 1 μg of GAL4-TK-CAT reporter and 0, 0.5, 1, or 2 μg of PM2-KLIP1 plasmid DNA. Percentage denotes the ratio between the acetylated substrate and the whole amount of substrate used in the assay. Results are the averages and standard deviations from three independent experiments except in panels A and F, which are the averages and the high and low values from two independent experiments.