Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Cyclic AMP-dependent protein kinase A and EPAC mediate VIP and secretin stimulation of PAK4 and activation of Na + ,K + -ATPase in pancreatic acinar cells

doi: 10.1152/ajpgi.00275.2018

Figure Lengend Snippet: Effect of ESI-09 and HJC0197, EPAC inhibitors, and KT-5720 and PKI, PKA inhibitors, on PAK4 activation (A), CREB phosphorylation (B), p44/42 phosphorylation (C), and p38 phosphorylation (D) by secretin in rat pancreatic acini. Isolated pancreatic acini were incubated in the absence or presence of ESI-09 (100 µM), HJC0197 (10 µM, 2h), KT-5720 (10 µM), or PKI (10 µM) for 1 h and then incubated with no addition (control) or secretin (10 nM) for 15 min, and then lysed. Western blots were analyzed using anti-pS474 PAK4, pS133 CREB, pT202/Y204 p44/42, or pT180/Y182 p38 and, as loading control, antitotal PAK4, antitotal CREB, antitotal p44/42, and antitotal p38. Bands were visualized using chemiluminescence and quantified by densitometry. Top: results of a representative blot of 3 independent experiments are shown. Bottom: means ± SE of at least 4 independent experiments. Results are expressed as percentages of basal stimulation of the control group. *P < 0.05 compared with the control group; $P < 0.05 compared with secretin alone. PAK, p21-activated kinase; PKI, PKA inhibitor.

Article Snippet: KT-5720 was from Abcam (Cambridge, MA).

Techniques: Activation Assay, Isolation, Incubation, Western Blot

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Cyclic AMP-dependent protein kinase A and EPAC mediate VIP and secretin stimulation of PAK4 and activation of Na + ,K + -ATPase in pancreatic acinar cells

doi: 10.1152/ajpgi.00275.2018

Figure Lengend Snippet: Effect of ESI-09 and HJC0197, EPAC inhibitors, and KT-5720 and PKI, a PKA inhibitors, on PAK4 activation (A), CREB phosphorylation (B), p44/42 phosphorylation (C), and p38 phosphorylation (D) by VIP in rat pancreatic acini. Isolated pancreatic acini were incubated in the absence or presence of ESI-09 (100 µM), HJC0197 (10 µM, 2 h), KT-5720 (10 µM) or PKI (10 µM) for 1 h and then incubated with no addition (control) or VIP (10 nM) for 15 min, and then lysed. Western blots were analyzed using anti-pS474 PAK4, pS133 CREB, pT202/Y204 p44/42 or pT180/Y182 p38 and, as loading control, antitotal PAK4, antitotal CREB, antitotal p44/42, and antitotal p38. Bands were visualized using chemiluminescence and quantified by densitometry. Top: results of a representative blot of 3 independent experiments are shown. Bottom: means ± SE of at least 4 independent experiments. Results are expressed as percentages of basal stimulation of the control group. *P < 0.05 compared with the control group; $P < 0.05 compared with VIP alone. PAK, p21-activated kinase; PKI, PKA inhibitor; VIP, vasoactive intestinal polypeptide.

Article Snippet: KT-5720 was from Abcam (Cambridge, MA).

Techniques: Activation Assay, Isolation, Incubation, Western Blot