Journal: Journal of Lipid Research

Article Title: Identification of miR-185 as a regulator of de novo cholesterol biosynthesis and low density lipoprotein uptake

doi: 10.1194/jlr.M041335

Figure Lengend Snippet: miR-185 downregulates SREBP-2 mRNA and protein level by binding to MREs within SREBP-2 mRNA 3′UTR. A: Luciferase activity was quantitated in 293T cells transfected with a control luciferase reporter plasmid (Con Luc), SREBP-2 3′UTR containing reporter plasmid (3′UTR), SREBP-2 3′UTR mutant (3′UTR MUT), and pre-miR-185 (miR185) or control pre-miR (Con miR). Luciferase activity was measured in 293T cells as described in the Materials and Methods section. Bar graphs represent mean ± SEM from three independent experiments. B: The fold change in SREBP-2 mRNA was measured by quantitative real-time-PCR in pre-miR-185, Antagamer-miR-185 (Antagomer) (100 nM), or control miR (Con miR) transfected HepG2 cells. Bar graphs represent mean ± SEM from three independent experiments. C: SREBP-2 mRNA was measured by quantitative real-time-PCR in pre-miR-185 or control miR transfected THLE-2 cells. Values were normalized to the level of GAPDH. Bar graphs represent mean ± SEM from three independent experiments. D: SREBP-2 protein level was determined using whole cell lysates by Western analysis in miR-185 overexpressing and control miR transfected HepG2 cells. β-Actin was used as a loading control. FL, full-length SREBP-2; M, mature form of SREBP-2.

Article Snippet: THLE-2, HepG2, and 293T cells were obtained from ATCC.

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Control, Plasmid Preparation, Mutagenesis, Real-time Polymerase Chain Reaction, Western Blot

Journal:

Article Title: The Long Terminal Repeat of Jaagsiekte Sheep Retrovirus Is Preferentially Active in Differentiated Epithelial Cells of the Lungs

doi:

Figure Lengend Snippet: Cell lines used in this study

Article Snippet: H441 and H358 cells were grown in RPMI 1640 medium (Gibco BRL) adjusted to contain 1.5 g of sodium bicarbonate per liter, 4.5 g of glucose per liter, and 10 mM HEPES with 10% FBS. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Cell line Origin Tissue or cell type ATCC no. or reference A549 Human Lung carcinoma CCL-185 H441 Human BAC HTB-174 H358 Human BAC CRL-5807 293T Human Embryonic kidney Lebkowsky et al. ( 33 ) OA1 Sheep Brain fibroblast CRL-6538 OAT Sheep Sheep testis CRL-6546 FLL Sheep Primary fetal lamb MRI a JS7 Sheep BAC Jassim ( 30 ) CP-MRI Sheep Choroid plexus MRI a CP-ATCC Sheep Choroid plexus CRL-1700 mtCC1-2 Mouse Clara cell Magdaleno et al. ( 34 ) BV2 Mouse Microglia A. Tenner a F9 Mouse Testicular carcinoma CRL-1720 FOP Mouse Mammary carcinoma J. Hassel a IC-21 Mouse Peritoneal macrophage TIB-186 MHS Mouse Alveolar macrophage CRL-2019 C2C12 Mouse Myoblast CRL-1772 ABI-2 Mouse Hybridoma HB-33 MLE-12 Mouse Lung epithelium CRL-2110 TCMK Mouse Mouse kidney CCL-139 NIH 3T3 Mouse Mouse embryo CCL-92 MLE-15 Mouse Type II pneumocyte Wikenheiser et al. ( 59 ) ST3 Mouse Thymus stroma Brightman et al. ( 10 ) Open in a separate window a Cells were provided directly by an investigator or institution with no reference available.

Techniques: