Fig. 7
From: Acid ceramidase targeting pyruvate kinase affected trypsinogen activation in acute pancreatitis
SPH binding to pyruvate kinase regulated trypsinogen activation. Primary pancreatic acinar cells were treated with saline or 200 nM of cerulein plus 10 ng/mL of LPS for 6 h. Cells were lysed, and the lysate was incubated with biotin-conjugated sphingosine. Proteins binding to the sphingosine were enriched in streptavidin magnetic beads. The potential target identified by mass spectrometry is shown (A) and validated by Western blotting (B). C–G Primary cells were treated with solvent, 5 μM of sphingosine, 10 μM of DASA58 plus sphingosine, and 200 nM of cerulein plus 10 ng/mL of LPS and DASA58 with LPS and sphingosine for 6 h. Cells were lysed for trypsin (C, D) and cathepsin B (E, F) assay, or were stained with mitochondrial ROS-specific dye, and detected by FACS (G)