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: Cancer Science

Article Title: Tumor exosomal RNPEP promotes lung metastasis of liver cancer via inducing cancer‐associated fibroblast activation

doi: 10.1111/cas.16417

Figure Lengend Snippet: RNPEP was overexpressed in hepatocellular carcinoma (HCC) tumors and cell lines. (A) RNPEP expression levels in human cancers in accordance with the data from Timer 2.0. (B) RNPEP expression levels in normal, HCC tumor and HCC metastatic tissues according to the data from the TNM plot. (C) RNPEP mRNA levels in normal human liver epithelial cells (THLE‐2), low‐metastatic HCC cells (MHCC‐97L), and high‐metastatic HCC cells (HCC‐LM3 and MHCC‐97H). Data were acquired by the real‐time PCR analysis and presented as mean ± SD, n = 3/group, * p < 0.05, ** p < 0.01. (D) RNPEP concentrations in cell supernatant of THLE‐2, MHCC‐97L, HCC‐LM3 and MHCC‐97H. Data were acquired by ELISA, and presented as the mean ± SD, n = 3/group, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: A normal human liver epithelial cell line (THLE‐2) was purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China) and cultured in cell‐specific culture medium (CM‐0833, Procell Life Science & Technology) at 37°C in a 5% CO 2 in air incubator.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay