Anti 5 ht antibody

Floxed Stat5 mice were a generous gift from Dr Lothar Hennighausen (National Institute of Health, USA) (Lee et al. 2007) (link). Tamoxifen-inducible islet-specific Stat5 knockout mice were generated by crossing Pdx1-Cre-ER mice with floxed Stat5 mice (Lee et al. 2007) (link). Tamoxifen (75 mg/kg body weight) or corn oil was injected intraperitoneally into 6-week-old mice. These mice were mated at 12 weeks old, and sacrificed at G12.5. After perfusion with 4% (w/v) paraformaldehyde, pancreata were isolated from the mice and embedded in paraffin. Immunofluorescent staining was performed using an anti-5-HT antibody (1:1000, Immunostar, Hudson, WI, USA), an anti-Ki-67 antibody (1:200, BD Biosciences, Tokyo, Japan), and an anti-insulin antibody (1:1000 dilution, Dako, Tokyo, Japan) as described previously (Kim et al. 2010) (link). Primary antibodies were detected with Alexa-fluor 488-and 555conjugated secondary antibodies and visualized using TCS-SP5 (Leica, Tokyo, Japan) and the Leica application Suite Version 2.6.0 software. Animal experiments were performed with the approval of the Ethics Review Committee for Animal Experimentation of Juntendo University and Korea Advanced Institute of Science and Technology.