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. Author manuscript; available in PMC: 2012 Nov 1.
Published in final edited form as: FEBS J. 2011 Oct 17;278(22):4338–4350. doi: 10.1111/j.1742-4658.2011.08351.x

Figure 3.

Figure 3

Effect of plasmid concentration on CTRC secretion from transfected HEK 293T cells. Transfections were performed with increasing concentrations of wild-type and N52S CTRC expression plasmids (from 25 ng to 1000 ng). Total plasmid DNA in the transfections was kept constant at 1000 ng by adding empty pcDNA3.1(−) vector. A. Conditioned media were analyzed by 12% SDS-PAGE and Coomassie blue staining. B. CTRC activity in the conditioned medium was measured with the Suc-Ala-Ala-Pro-Phe-p-nitroanilide substrate as described in Experimental Procedures. Activity was expressed as percentage of CTRC activity measured in the conditioned media of cells transfected with 1000 ng wild-type CTRC plasmid.