Journal: International Journal of Clinical and Experimental Pathology

Article Title: MiR-16 modulate temozolomide resistance by regulating BCL-2 in human glioma cells

doi:

Figure Lengend Snippet: Correlation between miR-16 expression and temozolomide resistance in glioma cells. A: The expression of miR-16 in different glioma cell lines by qRT-PCR. B: The correlation between the relative miR-16 expression and the IC50 values in glioma cells was quantified by Spearman’s rank correlation. C: Temozolomide-sensitive AM38 cells were transfected with specific inhibitor to miR-16. D: Temozolomide-resistant U251MG/TR cells were transfected with miR-16 mimics. After incubation with temozolomide for 48 hr, cell viability was assessed using CCK-8 assay and IC50 value to temozolomide was calculated. All experiments were performed in triplicate.

Article Snippet: Then, quantitative real-time RT-PCR (qRT-PCR) was performed using One Step SYBR ® PrimeScript™ RT-PCR Kit II (TaKaRa, China) according to standard protocol.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Incubation, CCK-8 Assay

Journal: Virulence

Article Title: gga-miR-1603 and gga-miR-1794 directly target viral L gene and function as a broad-spectrum antiviral factor against NDV replication

doi: 10.1080/21505594.2020.1864136

Figure Lengend Snippet: Detection of miRNA oligonucleotide efficiency by qRT-PCR assay . The indicated miRNA oligonucleotides (100 nM) were transfected into DF-1 cells for 24 h and untreated DF-1 cells were used as a blank control. The expression level of miRNAs was measured by qRT-PCR and normalized to 5S rRNA levels

Article Snippet: Viral L gene expression in RNA extractions was measured by qRT-PCR using TransScript Green One-Step qRT-PCR Super Mix (TransGen Biotech, China) on a LightCycler 480 machine (Roche, Switzerland).

Techniques: Quantitative RT-PCR, Transfection, Expressing

Journal: Virulence

Article Title: gga-miR-1603 and gga-miR-1794 directly target viral L gene and function as a broad-spectrum antiviral factor against NDV replication

doi: 10.1080/21505594.2020.1864136

Figure Lengend Snippet: Gga-miR-1603 and gga-miR-1794 negatively regulate viral L gene expression at both the protein and RNA levels . (a, c) DF-1 cells were co-transfected with pCMV-Flag-La L (a) or pCMV-Flag-ZJ1 L (c) and indicated miRNA oligonucleotides. After 24 h, total protein in the cells was harvested for western blotting. The relative expression of L protein was analyzed using Image J software, and this is shown in (b) for the results of (A) and (d) for (C) . (e, f) DF-1 cells were transfected with indicated miRNA oligonucleotides of gga-miR-1603 (e) and gga-miR-1794 (f) for 24 h and then the cells were inoculated with different NDV isolates (0.1 MOI). Total RNA was extracted for qRT-PCR at 24 hpi. Viral L gene expression was normalized to GAPDH using the 2 −ΔΔCt method. * P < 0.05 and ** P < 0.01

Article Snippet: Viral L gene expression in RNA extractions was measured by qRT-PCR using TransScript Green One-Step qRT-PCR Super Mix (TransGen Biotech, China) on a LightCycler 480 machine (Roche, Switzerland).

Techniques: Expressing, Transfection, Western Blot, Software, Quantitative RT-PCR

Journal: Virulence

Article Title: gga-miR-1603 and gga-miR-1794 directly target viral L gene and function as a broad-spectrum antiviral factor against NDV replication

doi: 10.1080/21505594.2020.1864136

Figure Lengend Snippet: Newcastle disease virus (NDV) infection exerts no effect on gga-miR-1603 and gga-miR-1794 expression in vitro . (a, b) DF-1 cells were inoculated with indicated NDV strains (0.1 MOI). The total miRNAs were extracted from cells collected at indicated time points post-NDV infection. Then, the expression levels of gga-miR-1603 (a) and gga-miR-1794 (b) were detected by qRT-PCR. (C, E, F) Three different avian cells lines, including CEF (c), HD11 (d), and LMH (e), were infected with the La Sota strain (0.1 MOI). The expression level of both miRNAs at different time points was measured using qRT-PCR. The relative expression of miRNAs was calculated by normalizing levels to 5S rRNA expression

Article Snippet: Viral L gene expression in RNA extractions was measured by qRT-PCR using TransScript Green One-Step qRT-PCR Super Mix (TransGen Biotech, China) on a LightCycler 480 machine (Roche, Switzerland).

Techniques: Infection, Expressing, In Vitro, Quantitative RT-PCR

Journal: Infectious Agents and Cancer

Article Title: HIV Nef enhances the expression of oncogenic c-MYC and activation-induced cytidine deaminase in Burkitt lymphoma cells, promoting genomic instability

doi: 10.1186/s13027-020-00320-9

Figure Lengend Snippet: Fig. 1 AID and c-MYC expressions are altered in B cells upon exposure to HIV Nef. a. Bar graphs showing relative mRNA expression of c-MYC (left panel) and AICDA (right panel) at 3 h, 6 h and 12 h exposure to HIV Nef, in lymphoblastoid cells (LCLs) and Burkitt lymphoma cells (Ramos). Cells were treated at the indicated concentrations of recombinant Nef, total RNA was isolated and qPCR was performed on reverse transcribed mRNA using primers specific to c-MYC or AICDA. The levels were normalised to the internal control GAPDH and plotted relative to mock (0 ng/ml) treated cells. *** p ≤0.001; **p ≤0.01; * p ≤0.05. All experiments were performed in triplicate. Bars indicate standard deviation. b. Ramos cells were treated at the indicated concentrations of recombinant Nef, total protein was isolated, separated on 12% SDS-PAGE and analysed by western blotting with antibodies against c-MYC and AID. p38 was detected as a loading control. All experiments were performed in triplicate

Article Snippet: Real-time PCR was performed on a LightCycler® 480 (Roche, Germany) using KAPA SYBR® FAST qPCR Kit (Kapa Biosystems, South Africa).

Techniques: Expressing, Recombinant, Isolation, Reverse Transcription, Control, Standard Deviation, SDS Page, Western Blot