Fibroblasts were cultured from ear skin tissues of the ABCA12+/+, ABCA12+/Z9, and ABCA12Z9/Z9 pigs. Assays of lipid accumulation were performed according to instruction previously described (Smyth et al., 2008 ). Fibroblasts were incubated in serum-containing medium with TO-901317 (4 mM; Cayman Chemical) and AcLDL (10 mg/ml; Yeasen Biotech) for 18 hrs. Fibroblasts were washed with PBS, fixed with 4% formaldehyde, and then stained with Oil Red O (Sigma-Aldrich) working solution.
Acldl
Manufactured by Yeasen
AcLDL is a laboratory equipment product. It is used for the detection and measurement of acetylated low-density lipoprotein (AcLDL) in biological samples.
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Lab products found in correlation
3 protocols using acldl
1
Lipid Accumulation Assay in Fibroblasts
2
Hepatic Cytochrome P450 Induction Assay
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In the ICG uptake assay, bioprinted structures or cells were cultured for 48 hours in a medium supplemented with 8-bromo-cAMP, pregnenolone-16α-carbonitrile, and progesterone, as per the manufacturer's instructions. Subsequently, they were coincubated with ICG (1 mg/ml) at 37°C for 1 hour, washed three times with PBS, and then switched to HEM culture medium. The uptake/release of ICG was observed under a microscope. Bioprinted structures or cells were stained with ac-LDL (Yeasen) and periodic acid-Schiff (Solarbio), and the staining results were observed under a microscope. Different inducers were used to treat 3DP-liver and eHep cells for 48 hours, and the gene expression levels of Cyp1a2 (3-methylcholanthracene, 10 μM, Sigma-Aldrich), Cyp3a11 (rifampicin, 25 μM, Solarbio), Cyp2e1 (rifampicin, 25 μM, Solarbio and ethyl alcohol; 0.5%, v/v), Cyp3a41b (3-methylcholanthracene, 10 μM, Sigma-Aldrich), and Cyp1a1 (rifampicin, 25 μM, Solarbio and barbituric acid, 10 μM, Aladdin) were detected. The P450-Glo Cyp1a2 Induction/Inhibition Assay (Promega) was used, following the manufacturer's instructions, to detect Cyp1a2 enzyme activity in different states.
3
Acetylated LDL Uptake Assay
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hESC (D0) and VOrs (D4-D40) were washed with PBS three times and then incubated with 10 μg/ml Ac-LDL (Yeasen) in MV2 medium for 4 hr at 37°C. The samples were washed three times with PBS, before confocal imaging using ×20 objective lens.
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