Figure 5.

ETS1 upregulates the expression of CEACAM1.

(A) CEACAM1 promoter was cloned upstream to firefly luciferase and co-transfected with a normalizing construct of Renilla luciferase, together with a vector encoding for ETS1 (ETS1), mutated ETS1 (ETS1-T38A), or an empty vector (Mock). Relative promoter activity was calculated relative to the effect of transfection with Mock vector. (B) Wild type (WT) or deletions in the putative ETS1 binding site in the negative strand (delETS1(−)), positive strand (delETS1(+)), or both (double) of the CEACAM1 promoter were cloned upstream to firefly luciferase and co-transfected with a normalizing construct of Renilla luciferase, together with a vector encoding for ETS1 (ETS1) or an empty vector (Mock). Data shown are normalized to Mock. (C) The effect of ETS1 (ETS1) compared to an empty vector (Mock) on CEACAM1 isoform (long, short) expression following transfected into melanoma was tested at the mRNA level using RT-PCR. (D) CEACAM1 expression was tested at the protein level using flow cytometry. The histograms of each of the transfectants are indicated in the figure. (A-C) The average results of three independent experiments. Significance was tested with Student’s t test, ** depicts P value of <.01.