Liver and placenta tissues were fixed in 4% paraformaldehyde, dehydrated and embedded in paraffin. The tissues were serially sectioned into 4 μm sections, and stained with hematoxylin and eosin. All sections were mounted onto a glass slide and observed under the light microscope and collected by NIS-Elements 3.2 (Nikon, Tokyo, Japan). Sections were examined by a qualified and blinded pathologist to evaluate the pathological changes.
Tissue sections were deparaffinized and rehydrated using ethanol and distilled water, and treated with 3% H2O2. Sections were then rinsed twice and incubated with goat serum to block non-specific antibody binding. Immunohistochemistry was performed using the primary antibodies for Cyp7a1 (Cat#bs-21430R, Bioss, 1:100), Cyp8b1 (Cat#bs-14165R, Bioss, 1:100), Cyp27a1 (Cat#bs-5049R, Bioss, 1:100), MRP2 (Cat#bs-1092R, Bioss, 1:100) and BSEP (Cat#bs-12440R, Bioss, 1:100). After washing, sections were incubated with the secondary antibody (PV-6001; Zhongshan, China) for 30 min. The sections were stained with DAB, dehydrated with ethanol and xylene, and then sealed. The slides were photographed using a digital microscope camera and collected by NIS-Elements 3.2 (Nikon, Tokyo, Japan).
Tissue sections were deparaffinized and rehydrated using ethanol and distilled water, and treated with 3% H2O2. Sections were then rinsed twice and incubated with goat serum to block non-specific antibody binding. Immunohistochemistry was performed using the primary antibodies for Cyp7a1 (Cat#bs-21430R, Bioss, 1:100), Cyp8b1 (Cat#bs-14165R, Bioss, 1:100), Cyp27a1 (Cat#bs-5049R, Bioss, 1:100), MRP2 (Cat#bs-1092R, Bioss, 1:100) and BSEP (Cat#bs-12440R, Bioss, 1:100). After washing, sections were incubated with the secondary antibody (PV-6001; Zhongshan, China) for 30 min. The sections were stained with DAB, dehydrated with ethanol and xylene, and then sealed. The slides were photographed using a digital microscope camera and collected by NIS-Elements 3.2 (Nikon, Tokyo, Japan).
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