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. 2008 Jul 30;105(31):11026–11031. doi: 10.1073/pnas.0805496105

Fig. 5.

Fig. 5.

GPC-PDE activity of GDPD5 immuno-precipitated from HEK293 cells and assayed in vitro. HEK293 cells were transfected for 24 h at 300 mosmol/kg with empty vector-V5 or GDPD5-V5. GPC-PDE activity was measured in immuno-precipitates from cell extracts and is normalized to the amount of extracted protein used for the immuno-precipitation. GDPD5-V5 expression was measured in the immuno-precipitates. Top panels are representative Western blots showing equivalent expression of GDPD5-V5 protein. (A) GPC-PDE activity is present in immuno-precipitates of GDPD5-V5, but not of empty vector-V5. (B) GDPD5 was immuno-precipitated from cells at 300 mosmol/kg, and its GPC-PDE activity measured in vitro, using the default buffer or buffers to which 200 mosmol/kg of NaCl, KCl, or urea was added. (C) After transfection, cells were incubated for 1 h either at 300 mosmol/kg or at 500 mosmol/kg (200-mosmol/kg NaCl or urea added). Note that GPC-PDE activity measured under identical conditions in vitro varies with the osmolality to which the cells had been exposed. (Mean ± SEM, n = 3, *, P < 0.05).