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. Author manuscript; available in PMC: 2010 Sep 1.
Published in final edited form as: Gut. 2009 Nov 30;59(3):365–372. doi: 10.1136/gut.2009.198903

Figure 2.

Figure 2

Diminished secretion and intracellular retention/degradation of the p.A73T CTRC mutant. Western blot analysis of conditioned media (A) and cell lysates (B) of pancreatic acinar cells expressing wild type CTRC, enhanced green fluorescent protein (control) or the p.A73T CTRC mutant. Twenty-four hours after transfection cells were washed twice with phosphate buffered saline and fresh OptiMEM medium was added. After 15 min incubation at 37 °C, the medium was collected (“basal”) and fresh OptiMEM containing 10 nM (AR42J) or 100 pM (mouse acini) concentration of cerulein was added to stimulate secretion. The “cerulein” medium was collected after 15 min incubation at 37 °C, and cells were also harvested for western blot analysis. Aliquots of conditioned media (100 µL) and cell lysates (20 µg total protein) were loaded onto a 15% SDS-polyacrylamide gel, transferred to Immobilon-P membrane and CTRC was detected with an antibody against the Glu-Glu epitope tag. Representative blots of three independent experiments are shown.