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. 2006 Nov 13;27(4):1348–1355. doi: 10.1128/MCB.00658-06

FIG. 3.

FIG. 3.

SYT located at the promoter region of COM1. (A) Luciferase activity of deletion mutants of the COM1 promoter in the presence of SYT-SSX1 protein. pB1 (kb −4 ∼ +22), pB2(kb −2.8 ∼ +22), or pB3(kb −1.5 ∼ +22) was the pGL3-basic reporter plasmid containing deletion mutants (the 4-kbp, 2.8-kbp, or 1.5-kbp upstream region, respectively, from the transcriptional start site) of the COM1 promoter. pP4 (kb −4 ∼ −1.5) was the pGL3 promoter vector with an SV40 promoter containing the region from kb 1.5 to 4 of the COM1 promoter. These constructs (100 ng) were cotransfected with (500 ng) or without SYT-SSX1 into NIH 3T3 cells. The activity for each promoter in the absence of SYT-SSX1 was set at 1. (B) Binding activity of SYT-SSX1 or SYT to the 1.5-kbp upstream region from the transcriptional start site of the COM1 promoter. HEK293T cells were transfected with the vector alone (5 μg), HA-tagged SYT-SSX1 (5 μg), or HA-tagged SYT (5 μg). After 3 days of incubation, the cells were treated with 1% formaldehyde to cross-link proteins to DNA and processed by ChIP assay with anti-HA antibody or immunoglobulin G (IgG) (negative control). DNA was amplified by PCR performed for 35 cycles.