Fig. 1.

Detection of Ano1 in pancreas and pancreatic islets. a RT-PCR of cDNA prepared from mRNA extracted from rat and human tissues. Transcripts of the expected size for Ano1 are observed (rat: 223 bp; human 314 bp). The 300-bp band is shown in the molecular weight marker column (MWM). Positive control: kidney. Negative control (Neg. control): no DNA. The sequencing of PCR products confirmed 100 % identity with the reference sequence for rat Ano1 cDNA complementary of rat Ano1 mRNA. b Western blot of Ano1 in rat islets, from left to right: molecular weight column (MW) showing the 100-, 150-, and 250-kDa bands, 80 μg rat islet lysate, 30 μg rat islet lysate, and 30 μg human thyroid lysate (positive control). Ano1 is detected at 119 kDa. c Immunofluorescence staining of pancreas section. c1 Immunohistochemical labeling (green-fluorescent Tyramide Alexa 488) of Ano1 in a section photomicrograph of rat pancreas. Most of the islet cells and acinar cells (at the level of apical pole) are labeled. c2 Counterstaining labeling by hematoxylin–eosin performed on the slice used for c1. c3 Specificity control: immunohistochemical labeling of Ano1 in a section photomicrograph of rat pancreas. The primary goat Ano1 antibodies (sc-69343) were coincubated in the presence of Ano1 synthetic peptide (ab97423) in a ratio 1:8. The labeling disappears. c4 Counterstaining labeling by hematoxylin–eosin performed on the slice used for c3. Arrows show islets. Scale bar is 50 μm