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. 2015 Nov 16;10(11):e0142991. doi: 10.1371/journal.pone.0142991

Fig 1. Direct regulation of the FZD10 gene by the SS18-SSX2 fusion protein.

Fig 1

A) Promoter activity in the regulatory region of the FZD10 gene. The upper panel indicated the 5’ region of the FZD 10 gene with amplified regions in the ChIP-qPCR analysis. The number indicates relative to the transcription start site, and the positions of the amplified region are: -1206 to -955 bp; -825 to -569 bp; -93 to +47 bp; +621 to +869 bp. The lower panel showed the promoter activities of fragments derived from this 5’ region in SYO-1. Error bars reflect SD in 3 experiments. B) Induction of SS18-SSX2 in U2OS. U2OS was transfected with an empty vector (EV) or 3xHA-tagged SS18-SSX2, and 48 h after the transfection, the expression of SS18-SSX2 was analyzed by Western blotting. The SS18-SSX2 and SS18 proteins were detected by an anti-SS18 antibody (top panel), and the 3xHA-SS18-SSX2 protein was detected by an anti-HA antibody (middle panel). C) Induction of FZD10 expression by SS18-SSX2 in U2OS. The expression of FZD10 was analyzed by RT-qPCR. Expression levels were normalized to those of human ACTB and calculated as fold changes relative to SYO-1. Error bars reflect SD in 3 experiments. **, p<0.01 by the t-test. D) Binding of SS18-SSX2 to the FZD10 locus. A ChIP assay with an anti-HA antibody and RT-qPCR were performed. The values indicate relative to rabbit IgG. Error bars reflect SD in 3 experiments.