Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: Pancreatic acinar-specific overexpression of Reg2 gene offered no protection against either experimental diabetes or pancreatitis in mice.

doi: 10.1152/ajpgi.00500.2009

Figure Lengend Snippet: Fig. 1. Pancreatic acinar-specific Reg2 overexpression. A: diagram of the Ela-Reg2 transgene. A 530-bp rat elastase I (Ela) promoter (13) was used to drive expression of a 0.7-kb mouse Reg2 cDNA (29) in the pcDNA3.1() vector (9). Arrows mark region covered by PCR primers ElaF and Reg2R for genotyping. B: representative PCR used for genotyping mice. A 1.3-kb Ela-Reg2 product was amplified from tails of transgenic, but not wild-type (WT), mice. Endogenous Reg1 gene was amplified as a control for the quality of genomic DNA. mRNA and protein levels and immunohistochemistry are shown in Figs. 4F, 4G, 5E, and 5F.

Article Snippet: The membrane was then incubated with rat anti-mouse Reg2 (1:2,000 dilution; R & D Systems, Minneapolis, MN) or rabbit anti- -actin (1:10,000 dilution; Sigma, St. Louis, MO) overnight at 4°C, washed with TBST, and incubated with horseradish peroxidase-conjugated secondary antibodies (Jackson Immunolabs, West Grove, PA) at room temperature for 1 h. The proteins were detected with enhanced chemiluminensce (GE Healthcare BioSciences, Piscataway, NJ), and the luminescent signal was captured using the Alpha Innotech FluorChem 8900 imaging system.

Techniques: Over Expression, Expressing, Plasmid Preparation, Transgenic Assay, Control, Immunohistochemistry

Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: Pancreatic acinar-specific overexpression of Reg2 gene offered no protection against either experimental diabetes or pancreatitis in mice.

doi: 10.1152/ajpgi.00500.2009

Figure Lengend Snippet: Fig. 5. Acinar expression of Reg2 in caerulein-induced acute pancreatitis. A: male Ela-Reg2 mice and wild-type littermates, 1.5–2 mo-old, were fasted for 16 h and then given 7 hourly injections of caerulein (50 g/kg ip). Mice were killed 11 h after the initial injection, and serum amylase level was measured (n 4–5). Values are means SE. **P 0.01 vs. saline-treated controls. B and C: hematoxylin-eosin (B) and amylase (C) staining of paraffin-embedded pancreatic tissues. Black, red, and white arrows indicate infiltrated neutrophils, apoptosis, and necrosis, respectively. Magnification 630. D: Western blot of poly(ADP-ribose)polymerase (PARP), intact protein or cleaved, in pancreatic tissue. -Actin was used as an internal control. E: changes in Reg2 protein level in response to transgenic overexpression and caerulein treatment. Top: representative Western blots. Bottom: summary of densitometry from 4–5 mice in each group. *P 0.05, **P 0.01 vs. saline-treated wild-type mice. F: pancreatic immunohistochemistry against Reg2 in untreated or caerulein-treated Ela-Reg2 and wild-type mice. Reg2 is revealed as dark brown patches.

Article Snippet: The membrane was then incubated with rat anti-mouse Reg2 (1:2,000 dilution; R & D Systems, Minneapolis, MN) or rabbit anti- -actin (1:10,000 dilution; Sigma, St. Louis, MO) overnight at 4°C, washed with TBST, and incubated with horseradish peroxidase-conjugated secondary antibodies (Jackson Immunolabs, West Grove, PA) at room temperature for 1 h. The proteins were detected with enhanced chemiluminensce (GE Healthcare BioSciences, Piscataway, NJ), and the luminescent signal was captured using the Alpha Innotech FluorChem 8900 imaging system.

Techniques: Expressing, Injection, Saline, Staining, Western Blot, Control, Transgenic Assay, Over Expression, Immunohistochemistry