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. 2013 Jan 7;8(1):e53453. doi: 10.1371/journal.pone.0053453

Figure 5. Murine M89V Siae is functionally normal.

Figure 5

Murine Siae cDNAs encoding WT, S127A, M89V, C196F and Q335P (murine equivalent of human Q309P SIAE) were introduced transiently into HEK 293T cells. Half of each lysate was immunoprecipitated with anti-Flag antibodies revealed in a Western blot assay (A, Lysate). The culture supernatants were also subjected to immunoprecipitation with anti-Flag antibodies to reveal the extent of secretion of each Siae variant at steady state (A, Supernatant). The other half of the cell lysate was similarly immunoprecipitated and examined for esterase activity, presented following normalization for lysate SIAE protein content (B). Each mutant was separately transfected and analyzed on three occasions. A representative experiment is shown. Error bars reflect esterase assays performed in triplicate in a single experiment.