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. 2014 May 5;9(5):e96610. doi: 10.1371/journal.pone.0096610

Figure 1. c-Jun could transactivate differentiation-associated genes.

Figure 1

(A) A schematic illustration of human cystatin A, involucrin and SPRR3 genes. AP-1 DNA binding sites predicted using bioinformatics software. (B) Down-regulation of c-Jun was frequently detected in the paired ESCC tissues (T) and adjacent nontumor tissues (N) from the same patient by Western blot (left panel). Statistical analysis was performed (right panel). (C) Expression of c-Jun was detected in esophageal cancer cell lines by Western blot. (D) c-Jun can directly bind to the promoters of these genes in vivo. KYSE450 cells were subjected to ChIP with anti-c-Jun or β-actin antibody. Purified DNA was analyzed by PCR. (E) AP-1 transactivation activity was detected by luciferase reporter assay system. KYSE450 cells were transiently cotransfected with pAP-1-luc and pRL-CMV. (F) c-Jun could transactivate differentiation- associated genes. KYSE450 cells were cotransfected with c-Jun and cystatin A, involucrin or SPRR3 promoter constructs. pRL-SV40 Renilla was used for normalization of transfection efficiency. After 48 hours, the luciferase activity was measured. Data shown are Mean ± SD from multiple independent experiments (*: P< 0.05).