Fig 5. Effect of UCN2 on NF-κB and trypsin activity and IκB degradation.

Rats were pre-treated with saline or UCN2 for 30 min followed by treatment with caerulein for 1 hour. (A) NF-κB DNA binding activities were measured in the pancreatic nuclear extracts using ELISA. UCN2 treatment decreased NF-κB by nearly 42% in the nuclear extracts. (B) Western blot analysis showed decreased IκB-α and NF-κB levels in pancreatic tissue cytosolic extract after caerulein treatment compared with saline controls, suggesting either increased degradation IκB-α or increased translocation of NF-κB from the cytosol to the nucleus. UCN2 treatment prevented caerulein-induced changes in IκB-α degradation and NF-κB nuclear translocation. GAPDH, a housekeeping protein was used as a loading control (n = 3/condition). (C) Trypsin activity was increased after caerulein treatment and UCN2 treatment had a modest (15%) effect on trypsin activity. Values are means ± SEM (n = 5). One-way ANOVA and Tukey’s multiple comparisons. CR: caerulein.