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. 2015 Dec 29;27(3):255–266. doi: 10.1089/hum.2015.110

Figure 2.

Figure 2.

Enhancer-blocking activity of candidate insulators. (a) Enhancer-blocking plasmid construct. Insulators or an FV vector LTR containing an insulator to be tested were placed in the multiple cloning site between the CMV enhancer and the CMV minimal promoter controlling the expression of mCherry. The ratio of mCherry expression to EGFP expression was then determined. SVPA, simian virus 40 (SV40) polyadenylation site; TKPA, thymidine kinase polyadenylation site. (b) Insulator activity. Enhancer-blocking plasmid with no insulator (Control) or with the indicated insulators was transfected into HEK293T cells and the effect of insulators on enhancers, determined by intensity of fluorescence, was evaluated by flow cytometry 40–45 hr posttransfection. Columns represent the percentage of normalized mCherry expression as compared with the uninsulated control. (c) Enhancer-blocking capacity of insulators in the context of the FV LTR. *p < 0.05 compared with control; **p < 0.05 between samples; #p < 0.05 compared with insulator alone.