Fig. 3.

Decreased marker signals in parotid glands of diabetic NOD mice. Immunohistochemistry of parotid glands from diabetic NOD mouse, non-diabetic NOD mouse and control mouse was performed with antibodies for anti-aquaporin 8 (A, upper panel). Distribution of aquaporin 8 (green) was shown with that of aquaporin 5 (red), as a marker for acinar cells [16]. Phalloidin staining is shown in the lower images; white lines indicate cell edges. Arrows indicate lumina of acinar cells. Homogenates were prepared from parotid glands of diabetic NOD mouse, non-diabetic NOD mouse and control mouse (C57BL/6), and analyzed by immunoblotting for myoepithelial cell markers; anti-actin-α1, anti-actin-α2, and anti-aquaporin 8 (B). Aliquots of homogenates (10 μg) were applied on a polyacrylamide gel plate. Actin-α1 was detected in control mouse parotid glands but rarely in diabetic NOD mouse parotid glands. RT-PCR was performed with 2.5 ng/μl template total RNA, prepared from parotid glands according to methods described under “Section 2” (C). Amplicons were amplified over 35 cycles. Actb (β-actin) was used as a positive internal standard. Data show a representative result from three repeat experiments in four animals. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)