Figure 2 | Scientific Reports

Figure 2

From: Regulatory elements in vectors containing the ctEF-1α first intron and double enhancers for an efficient recombinant protein expression system

Figure 2

Construction of cassettes with/without the wild-type (wt) EF-1α first intron containing two translational enhancer sequences (CMV and SV40) downstream of SV40 polyA. (a) GFP expression after transfection of HEK293T cells with the two indicated constructs that included two enhancers and the presence or absence of the wtEF-1α first intron cassette. Protein expression analyses using fluorescence microscopy and quantification of the fluorescence image density. (b) Schematic representation of wt, double C-terminal (ct) and ct only EF-1α first intron cassettes with two translational enhancer sequences downstream of SV40 polyA. (c) GFP mRNA levels in HEK293T cells assessed using a real-time PCR assay. The gene expression levels, normalized to β-actin expression, are expressed as relative change, setting the values of the duodenum as one. The results are presented as the mean ± SD (n = 3 experiments). (d) Immunoblot analysis of GFP protein levels. (e) The immunoblot bands were quantified by densitometry analysis, and the β-actin ratios were calculated.

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