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: Biomedicines

Article Title: Impaired Function of Solute Carrier Family 19 Leads to Low Folate Levels and Lipid Droplet Accumulation in Hepatocytes

doi: 10.3390/biomedicines11020337

Figure Lengend Snippet: Lipid droplet accumulation in SLC19A1 -knockdown THLE2-cells. SLC19A1 -KD THLE2-cells develop steatosis spontaneously and show impaired lipid metabolism. DAPI (blue staining) for nucleic acids and LipoTox (green staining) for lipids. Scale bar added to 20× and 400× images. *: Difference between shRNA-Control and shRNA- SLC19A1 is statistically significant, p value<0.05.

Article Snippet: THLE2 human liver cell line (ATCC ® CRL-10149 TM ) [ ] was purchased from ATCC ® .

Techniques: Knockdown, Staining, shRNA, Control

Journal: Biomedicines

Article Title: Impaired Function of Solute Carrier Family 19 Leads to Low Folate Levels and Lipid Droplet Accumulation in Hepatocytes

doi: 10.3390/biomedicines11020337

Figure Lengend Snippet: THLE2 human liver cells cultured on folate-free RPMI supplemented with 10% dialyzed FBS. (A) Graphical representation of the fluorescence mean calculated in three wells per each treatment and three independent fields per well. *: Difference between fluorescence on “no supplemented cells” vs “cells supplemented with FA and/or 5MTHF” is statistically significant, p value < 0.05. ( B ) Images of the different culture conditions stained with LipidTOX TM (green) and DAPI (blue). Scale bar = 100 uM (10× objective).

Article Snippet: THLE2 human liver cell line (ATCC ® CRL-10149 TM ) [ ] was purchased from ATCC ® .

Techniques: Cell Culture, Fluorescence, Staining

Journal: Biomedicines

Article Title: Impaired Function of Solute Carrier Family 19 Leads to Low Folate Levels and Lipid Droplet Accumulation in Hepatocytes

doi: 10.3390/biomedicines11020337

Figure Lengend Snippet: Metabolomics fingerprint of SLC19A1 -KD cells. Heatmap represents metabolomic signatures of the comparison of SLC19A1 -KD and shControl THLE2-cells. Metabolites present in the picture were ordered according to their carbon number and unsaturation degree. In the second column, grey lines correspond to significant fold-changes of individual metabolite levels, darker grey color stress higher significances (Student’s t -test p < 0.05, p < 0.01, or p < 0.001).

Article Snippet: THLE2 human liver cell line (ATCC ® CRL-10149 TM ) [ ] was purchased from ATCC ® .

Techniques: Comparison