the hbv replication plasmid phbv1.3 which contained 1.3 copies of the hbv genome  (BioVector Inc)

Journal: Journal of Translational Medicine

Article Title: Upregulation of microRNA-328-3p by hepatitis B virus contributes to THLE-2 cell injury by downregulating FOXO4

doi: 10.1186/s12967-020-02299-8

Figure Lengend Snippet: Effect of HBV/HBs/HBc/HBp/HBx on pro-inflammatory cytokines, miR-328-3p expression, STAT3 phosphorylation, and FOXO4 protein expression. THLE-2 cells were transfected with pHBV1.3 (HBV replication plasmid), pHBs (expressing HBsAg), pHBc (expressing HBcAg), pHBp (expressing DNA polymerase protein), pHBx (expressing HBx), and empty pcDNA3.1 vector (control), under LPS stimulation (1 μg/mL, 24 h). a ELISA was performed to detect levels of TNF-α, IL-6, IL-8, IL-12, and IL-18. b qRT-PCR was conducted to examine miR-328-3p expression. c Western blot was performed to measure protein expression of FOXO4 and STAT3, and phosphorylated (p)-STAT3. Values are presented as the mean ± SD (n = 3). *P < 0.05, **P < 0.01 vs. NC. # P < 0.05, ## P < 0.01 vs. LPS + vector

Article Snippet: The HBV replication plasmid pHBV1.3 which contained 1.3 copies of the HBV genome was obtained from BioVector (Beijing, China).

Techniques: Expressing, Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

Journal: Journal of Translational Medicine

Article Title: Upregulation of microRNA-328-3p by hepatitis B virus contributes to THLE-2 cell injury by downregulating FOXO4

doi: 10.1186/s12967-020-02299-8

Figure Lengend Snippet: STAT3 activates miR-328-3p transcription and mediates the HBV/HBc/HBx-induced upregulation of miR-328-3p. a THLE-2 cells were cultured with recombinant human IL-6 protein (200 ng/mL), S3I-201 (50 μM), or DMSO (vehicle control) for 24 h, followed by qRT-PCR analysis of miR-328-3p expression. b Resultant ChIP DNA was amplified by qRT-PCR. CHIP-qPCR assay confirmed the direct binding of STAT3 to the miR-328 promoter. IgG served as a negative control. IL-6 was used to activate STAT3. c – e THLE-2 cells were transfected with pHBV1.3, pHBc, pHBx and empty pcDNA3.1 vector (control), followed by S3I-201 (50 μM, 24 h) or not, under LPS stimulation (1 μg/mL, 24 h). Then c qRT-PCR was conducted to examine miR-328-3p expression. d ELISA was performed to detect levels of TNF-α, IL-6, IL-8, IL-12, and IL-18. e Western blot was performed to measure protein expression of STAT3, and p-STAT3. Values are presented as the mean ± SD (n = 3). *P < 0.05, **P < 0.01 vs. LPS + Vector. ## P < 0.01 vs. LPS + pHBV1.3. && P < 0.01 vs. LPS + pHBc. $$ P < 0.01 vs. LPS + pHBx

Article Snippet: The HBV replication plasmid pHBV1.3 which contained 1.3 copies of the HBV genome was obtained from BioVector (Beijing, China).

Techniques: Cell Culture, Recombinant, Quantitative RT-PCR, Expressing, Amplification, Binding Assay, Negative Control, Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Journal of Translational Medicine

Article Title: Upregulation of microRNA-328-3p by hepatitis B virus contributes to THLE-2 cell injury by downregulating FOXO4

doi: 10.1186/s12967-020-02299-8

Figure Lengend Snippet: miR-328-3p inhibitor suppresses pHBV1.3-induced HBV activity. THLE-2 cells were co-transfected with pHBV1.3 or empty pcDNA3.1 vector (control), and miR-328-3p inhibitor or inhibitor negative control (NC). a HBV RNA and b HBV DNA were qualified. c , d The OD 450 absorbance values were obtained by ELISA for determining levels of HBsAg and HBeAg in THLE-2 cell supernatants. Values are presented as the mean ± SD (n = 3). ## P < 0.01 vs. Vector + inhibitor NC. && P < 0.01 vs. inhibitor NC + pHBV1.3

Article Snippet: The HBV replication plasmid pHBV1.3 which contained 1.3 copies of the HBV genome was obtained from BioVector (Beijing, China).

Techniques: Activity Assay, Transfection, Plasmid Preparation, Negative Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of Translational Medicine

Article Title: Upregulation of microRNA-328-3p by hepatitis B virus contributes to THLE-2 cell injury by downregulating FOXO4

doi: 10.1186/s12967-020-02299-8

Figure Lengend Snippet: FOXO4 suppresses pHBV1.3-induced HBV activity. THLE-2 cells were transfected with pHBV1.3 and pcDNA3.1-FOXO4, both alone and in combination. a HBV RNA and b HBV DNA were qualified. c The OD 450 absorbance values were obtained by ELISA for determining levels of HBsAg and HBeAg in THLE-2 cell supernatants. Values are presented as the mean ± SD (n = 3). **P < 0.01 vs. Vector. ## P < 0.01 vs. pHBV1.3

Article Snippet: The HBV replication plasmid pHBV1.3 which contained 1.3 copies of the HBV genome was obtained from BioVector (Beijing, China).

Techniques: Activity Assay, Transfection, Enzyme-linked Immunosorbent Assay, Plasmid Preparation

Journal: Journal of Translational Medicine

Article Title: Upregulation of microRNA-328-3p by hepatitis B virus contributes to THLE-2 cell injury by downregulating FOXO4

doi: 10.1186/s12967-020-02299-8

Figure Lengend Snippet: FOXO4 overexpression abrogates the miR-328-3p-mediated cell injury. THLE-2 cells were co-transfected with pHBV1.3 or empty pcDNA3.1 vector (control), and miR-328-3p mimic or mimic negative control (NC), under LPS (1 μg/mL, 24 h) and HBV stimulation. a Cell proliferation at different time points was examined by MTT assay. b Cell apoptosis was examined by flow cytometry and the apoptosis rate was shown. c Levels of TNF-α, IL-6, IL-8, IL-12, and IL-18 were measured by ELISA. Values are presented as the mean ± SD (n = 3). *P < 0.05, **P < 0.01 vs. LPS + HBV + Vector + mimic NC. # P < 0.05, ## P < 0.01 vs. LPS + HBV + Vector + miR-328-3p mimic

Article Snippet: The HBV replication plasmid pHBV1.3 which contained 1.3 copies of the HBV genome was obtained from BioVector (Beijing, China).

Techniques: Over Expression, Transfection, Plasmid Preparation, Negative Control, MTT Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay