Journal: Frontiers in pharmacology

Article Title: LncRNA-uc002mbe.2 Interacting with hnRNPA2B1 Mediates AKT Deactivation and p21 Up-Regulation Induced by Trichostatin in Liver Cancer Cells.

doi: 10.3389/fphar.2017.00669

Figure Lengend Snippet: FIGURE 7 | uc002mbe.2 knockdown inhibits the in vivo sensitivity of HCC cells to TSA. (A) A representative image of an isolated tumor from nude mice subcutaneously inoculated with uc002mbe.2 shRNA-transfected Huh7 cells and shGFP-transfected cells after 14 days of TSA treatment. (B) Tumor growth curve. (C) Mean tumor weight. The data are presented as the mean ± SD of 8 nude mice. ∗p < 0.05. (D) Total RNA extracted from isolated tumor tissue was used to evaluate the efficiency of uc002mbe.2 knockdown by quantitative real-time PCR. (E) Total protein was isolated from xenograft tumors and subjected to Western blotting analyses to evaluate the levels of hnRNPA2B1, p-AKT and p21.

Article Snippet: Scramble siRNA and predesigned siRNA specific for human hnRNPA2B1 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States).

Techniques: Knockdown, In Vivo, Isolation, shRNA, Transfection, Real-time Polymerase Chain Reaction, Western Blot

Journal: Molecules and Cells

Article Title: Hepatitis B Virus X Protein Stimulates Virus Replication Via DNA Methylation of the C-1619 in Covalently Closed Circular DNA

doi: 10.14348/molcells.2018.0255

Figure Lengend Snippet: (A) HepG2 cells were transfected with 1.2-mer WT as described in in the presence of an increasing concentration of 5-Aza-2′dC. For the determination of viable cell number, MTT assay was performed as described in Methods (n = 4). Levels of extracellular HBV particles were determined by IP-coupled conventional PCR. (B) HepG2 cells were transfected with either 1.2-mer WT or its HBx-null derivative for 48 h in the presence or absence of 5 μM 5-Aza-2′dC. Levels of extracellular HBV particles were determined by HBsAg ELISA (n = 5). (C) Levels of intracellular HBV DNA and (D) extracellular virus particles from HepG2 cells prepared as in (B) were determined by IP-coupled real-time PCR (n = 6). (E) HepG2 cells were transfected with either 1.2-mer WT or its HBx-null derivative along with the indicated amount of DNMT1 shRNA plasmid or SC shRNA plasmid for 48 h, followed by Western blotting. (F) Levels of extracellular HBV particles released from HepG2 cells prepared in (E) were determined by IP-coupled real-time PCR (n = 6).

Article Snippet: Scrambled (SC) shRNA plasmid and NREBP shRNA plasmid was purchased from Santa Cruz Biotechnology.

Techniques: Transfection, Concentration Assay, MTT Assay, Enzyme-linked Immunosorbent Assay, Virus, Real-time Polymerase Chain Reaction, shRNA, Plasmid Preparation, Western Blot

Journal: Molecules and Cells

Article Title: Hepatitis B Virus X Protein Stimulates Virus Replication Via DNA Methylation of the C-1619 in Covalently Closed Circular DNA

doi: 10.14348/molcells.2018.0255

Figure Lengend Snippet: (A) Schematic representation of pHBV-luc and its derivatives. The basal luciferase activities of these constructs along with an empty vector pGL2-luc (Promega) were compared in HepG2 cells (n = 4). (B to G) Either an empty vector or HBx-expression plasmid was co-transfected with the indicated reporter construct into HepG2 cells for 48 h, followed by luciferase assay. The values indicate the relative luciferase activity compared to the basal level of the control (n = 4). (B, D, E, and G) Cells were either mock-treated or treated with 5 μM 5-Aza-2′dC. (C and F) The indicated amount of DNMT1 shRNA plasmid or SC shRNA plasmid was included in the transfection mixtures.

Article Snippet: Scrambled (SC) shRNA plasmid and NREBP shRNA plasmid was purchased from Santa Cruz Biotechnology.

Techniques: Luciferase, Construct, Plasmid Preparation, Expressing, Transfection, Activity Assay, Control, shRNA

Journal: Molecules and Cells

Article Title: Hepatitis B Virus X Protein Stimulates Virus Replication Via DNA Methylation of the C-1619 in Covalently Closed Circular DNA

doi: 10.14348/molcells.2018.0255

Figure Lengend Snippet: (A–C) HepG2 cells were co-transfected with pHBV-luc (A), pHBV-NRE-luc (B), or pHBV-Enh-luc (C) with either 1.2-mer WT or 1.2-mer HBx-null in the presence or absence of NREBP shRNA plasmid for 48 h, followed by luciferase assay (n = 5). (D) HepG2 cells were transfected with either 1.2-mer WT or 1.2-mer HBx-null with or without NREBP shRNA plasmid for 48 h. Levels of extracellular and intracellular HBV DNA and E-cadherin DNA as an internal control were determined by PCR (upper panels). Levels of HBx, HBc, and NREBP were determined by Western blotting (lower panels). (E) HepG2 cells were transfected with either 1.2-mer WT or 1.2-mer HBx-null for 48 h for 48 h in the presence or absence of 5 μM 5-Aza-2′dC. ChIP assay was performed to determine levels of NREBP bound on the NRE of HBV cccDNA (upper panels).

Article Snippet: Scrambled (SC) shRNA plasmid and NREBP shRNA plasmid was purchased from Santa Cruz Biotechnology.

Techniques: Transfection, shRNA, Plasmid Preparation, Luciferase, Control, Western Blot