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. 2016 Apr 13;6:190–196. doi: 10.1016/j.bbrep.2016.04.004

Fig. 1.

Fig. 1.

Expression of mouse N-DSP in mammalian and bacterial expression systems. (A, B) Expression of N-DSP in HEK293 cells. The mammalian expression vector encoding N-DSP (containing mouse N-terminal portion of DSP; 1–47 amino acids) fused to V5/His tag or an empty V5/His-tag vector (EV) was transiently transfected into HEK293 cells, the conditioned media or cell lysates were collected, immunoprecipitated (IP) with anti-V5 antibody and subjected to Western blot analysis (WB) with either anti-V5 (A) or anti-DSP (B) antibody. *; IgG light chain. (C) Expression of GST-N-DSP in bacterial expression system. The equal amount (500 ng each) of GST (lane 1) and GST-N-DSP proteins (lane 2) were separated by 4–12% SDS-PAGE and stained with CBB. The presence of a single band in each lane confirms the purity of GST proteins. The same proteins were also analyzed by Western blot analysis (lanes 3–6). The immunoreactivities to anti-GST antibody were clearly detected in both GST proteins (lanes 3 and 4). The immunoreactivity to anti-DSP antibody was detected only in GST-N-DSP protein (lane 6), but not in GST alone (lane 5), demonstrating the successful generation of GST-N-DSP protein.