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. 2014 May 13;5(10):3386–3398. doi: 10.18632/oncotarget.1960

Figure 5. Transcriptional regulation of neogenin-1 through interaction with galectin-3 and HSF-1 transcriptional factor.

Figure 5

(A) The proximal HSF-1 binding site is located between -1478 and -1470 from the initiation codon of NEO1 gene. (B) Chromatin immunoprecipitation assay was performed as described in “Methods.” Total genomic DNA was used in the input lane as a control for the PCR assay. (C) Luciferase activity of HSF-1 in AGS cells treated with galectin-3 siRNA or HSF-1 siRNA. A luciferase assay was performed using a HSF-1 binding site luciferase vector transfection with galectin-3 siRNA or HSF-1 siRNA (*, ** p<0.001). The experiments were performed in triplicate. (D) The detection of the interaction between galectin-3 and HSF-1 by immunoprecipitation was performed as described in “Methods,” and then galectin-3 and HSF-1 were detected by western blot analysis. Whole-cell lysate (WCL) was used as a loading control. (E) Protein levels of galectin-3 and HSF-1 were detected in the nuclear and cytosol fractions of AGS cells which were transfected with galectin-3 or HSF-1 siRNA. Lamin A/C was used as a control for the nuclear fraction.