Fig. 2.

Apical Na⁺-dependent bile acid transporter (ASBT) expression is increased in diabetes mellitus. Distal ileum was removed from control and diabetic animals, and mucosa was scraped as mentioned in materials and methods. Total RNA was extracted, and the relative expression of ASBT mRNA (A) and organic solute transporter (Ost) α and β mRNA (B) was assessed by real-time RT-PCR using SYBR green. The level of mRNA expression for each target gene was normalized to the expression of 18S mRNA in the same sample. In C, immunofluorescence staining for actin (red) in optimal cutting temperature (OCT)-embedded sections of rat ileum from control and diabetic rats showing long villi but no apparent change in the size of epithelial cells. In D, ileal segments were embedded in OCT media, sectioned to 9 μm thickness. The tissues were then mounted on special slides suitable for laser capture microdissection as mentioned in materials and methods. Ileal sections were stained with hematoxylin-eosin (H&E), and epithelial cells were visualized under the microscope. Equal number of cells was cut by laser from tissues from control and diabetic rats. Cells were collected, and total RNA was extracted. The relative expression of ASBT mRNA normalized to 18S mRNA was evaluated by real-time RT-PCR. Values represent means ± SE from at least 3–5 animals. *P ≤ 0.05 compared with control.