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. 2011 May 19;6(5):e19922. doi: 10.1371/journal.pone.0019922

Figure 2. Immunoblotting and enzyme activity measurement of feline PDE5 expressed in E. coli.

Figure 2

(a) Immunoblot analysis and coomassie blue staining of whole cell extracts made from E. coli cells carrying feline PDE5 expression plasmid pT7-PDE5 or a control vector pSNAP-tag(T7). Lanes 1 and 3 are extracts from cells carrying pSNAP-tag(T7). Lanes 2 and 4 are extracts from cells carrying pT7-PDE5. Lanes 1 and 2 are Coomassie blue staining of the gel. Lanes 3 and 4 are immunoblot using an anti-human PDE5 antibody. (b) Measurement of PDE5 activity. PDE activity of the extracts from cells carrying pT7-PDE5 in the presence or absence of a PDE5 inhibitor, MY5445, was shown (see Method for details). Basal level PDE activity determined with extract from cells carrying pSNAP-tag(T7) was subtracted. The error bars indicate standard deviation (n = 4).