Journal: PLoS ONE

Article Title: Lineage Diversion of T Cell Receptor Transgenic Thymocytes Revealed by Lineage Fate Mapping

doi: 10.1371/journal.pone.0001512

Figure Lengend Snippet: (A) Sox13 expression in male HY + DN thymocyte subpopulations, WT DN thymocyte subpopulations and TCRγδ + thymocyte subsets was determined by real time PCR. (B) Sox13 expression in peripheral T lymphocytes from WT and male HY transgenic mice. As a reference, Sox13 expression in TCRγδ + CD24 hi thymocytes, which was detected in the same real time PCR measurement, is shown together with Sox13 expression in peripheral lymphocyte populations. Relative Sox13 expression levels normalized against Hprt expression are shown. The data shown are representative from two experiments with similar results.

Article Snippet: The primer sequences for Sox13 were obtained from SuperArray (catalogue number PPM04782A-200).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Transgenic Assay

Journal: Oncotarget

Article Title: Hsa-miR-139-5p inhibits proliferation and causes apoptosis associated with down-regulation of c-Met

doi:

Figure Lengend Snippet: A549 and SK-MES-1 cells were transfected with miR-139-5p mimic and mimic NC, miR-139-5p inhibitor and inhibitor NC, for forty-eight hours, respectively. A. Shown are representative photomicrographs of A549 cells stained with Hoechst. Cells grown in coverslips were stained with Hoechst 33342 and photographed under a fluorescence microscope. Cells undergoing DNA fragmentation were counted manually. Bar = 50 μm. B. Quantitative representation of the number of apoptotic after transfecting cells with related miRNAs for forty-eight hours. The total number of cells (~200) with or without fragmented nuclei was counted, and the percentage of apoptotic cells was calculated. C. Shown are representative photomicrographs of SK-MES-1 cells stained with Hoechst. Cells grown in coverslips were stained with Hoechst 33342 and photographed under a fluorescence microscope. Cells undergoing DNA fragmentation were counted manually. Bar = 50 μm. D. Quantitative representation of the number of apoptotic after transfecting cells with related miRNAs for forty-eight hours. The total number of cells (~200) with or without fragmented nuclei was counted, and the percentage of apoptotic cells was calculated. E. Shown are representative photomicrographs of cells dual-stained with annexin-FITC/PI. Bar = 5 μm. F. Shown are representative photomicrographs of flow cytometric analysis. G. Statistical analysis of flow cytometric analysis. H. Expression of Bcl2 protein in transfected A549 cells. Western blot of Bcl2 protein in A549 cells after transfection. I. Upper, statistical analysis of Western blot; Lower, qRT-PCR of Bcl2 mRNA in A549 cells after transfection. J. Western blot of cleaved-caspase-3 and caspase-3 (total) protein in A549 cells after transfection. K. Statistical analysis of Western blot. L. Quantitative representation of caspase-3 activity in A549 Upper, and SK-MES-1 Lower, cells transfected with related miRNAs for forty-eight hours. Assays were performed in triplicate. Means ± SEM are shown. Statistical analysis was conducted using One-way ANOVA.

Article Snippet: For the qRT-PCR detection of mature miR-139-5p expression, we purchased the Bulge-LoopTM miRNA qRT-PCR Primer Set and the miRNA qRT-PCR Control Primer Set (both from RiboBio).

Techniques: Transfection, Staining, Fluorescence, Microscopy, Expressing, Western Blot, Quantitative RT-PCR, Activity Assay

Journal: Oncotarget

Article Title: Hsa-miR-139-5p inhibits proliferation and causes apoptosis associated with down-regulation of c-Met

doi:

Figure Lengend Snippet: Primer sequences for quantitative reverse transcription (RT)-PCR (miRNA)

Article Snippet: For the qRT-PCR detection of mature miR-139-5p expression, we purchased the Bulge-LoopTM miRNA qRT-PCR Primer Set and the miRNA qRT-PCR Control Primer Set (both from RiboBio).

Techniques: Reverse Transcription, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Evaluation of Various Alternative Economical and High Throughput SARS-CoV-2 Testing Methods within Resource-Limited Settings

doi: 10.3390/ijms232214350

Figure Lengend Snippet: Comparison of the sensitivity and specificity of the commercial SARS-CoV-2 RT-PCR assay kits.

Article Snippet: Five commercially available RT-PCR SARS-CoV-2 assay kits from different manufacturers were selected in this study for the method comparison including the Thermo Fisher TaqPathTM COVID-19 Assay Kit (Thermo Fischer Scientific, Pleasanton, CA, USA), an internationally approved kit, and the four alternative RT-PCR SARS-CoV-2 assay kits: Nucleic Acid COVID-19 Test Kit (SARS-CoV-2) (Wuhan Easy-diagnosis Biomedicine, Wuhan, China), abTES TM COVID-19 qPCR I Kit (Anatech Instrument (PTY) LTD, Meadowbrook, Business Estate, Sloane Park, Gauteng, South Africa), PCL COVID19 Speedy RT-PCR Kit (PCL Inc. Multiplex In Vitro Diagnostic Global Leader, Seoul, South Korea), and the PCLMD nCoV One-Step RT-PCR Kit (PCL Inc. Multiplex In Vitro Diagnostic Global Leader, Seoul, South Korea).

Techniques: Comparison, One Step RT-PCR

Journal: PLoS ONE

Article Title: Bacillus anthracis Peptidoglycan Stimulates an Inflammatory Response in Monocytes through the p38 Mitogen-Activated Protein Kinase Pathway

doi: 10.1371/journal.pone.0003706

Figure Lengend Snippet: (A) Time course of secreted TNFα induced by PGN in whole blood. PB was stimulated with PGN (10 µg/ml) and the supernatant was sampled at the indicated times. TNFα concentration was measured by ELISA. Data are expressed as mean±SEM for 3 replicates. Statistical analysis was by ANOVA with Bonferroni post test. *, p<0.001 versus 0 PGN; #, p<0.01 versus 0 PGN. (B) PB was stimulated with PGN (10 µg/ml)±SB202190 for 2 hours. RNA was isolated from leukocytes, converted to cDNA which was subjected to real-time quantitative PCR. Gene specific primers used were for TNFα and β-actin in a real-time PCR assay. Data presented are the mean±SEM of separate preparations of the TNFα/β-actin ratio from 4 individuals, 3 replicates each and normalized to the unstimulated ratio. Statistical analysis was by t test. *, p<0.01 for TNFα, PGN versus unstimulated; # p>0.3 for β-actin versus unstimulated. (C) PB was stimulated with PGN (10 µg/ml) for 3 hours. Cells were harvested to represent 100% RNA at 3 hours. To equivalent samples, actinomycin D was added immediately to block further RNA synthesis, or actinomycin+SB202190 were added to block RNA synthesis and the p38 MAP kinase pathway. Cells were harvested at 10, 20, 30, 60 minutes after these additions. RNA was isolated, cDNA was prepared and subjected to real-time quantitative PCR for TNFα as in (B). Quantity of RNA was expressed as % of RNA after 3 hours of stimulation. Data presented are the mean±SEM from 3 replicate PCR reactions from 1 individual. Statistical analysis was by ANOVA with Bonferroni post test of *, p<0.001 for PGN+ActD versus PGN+ActD+SB; #, p<0.01.

Article Snippet: Real-time quantitative PCR was performed on 20 μl volumes using RT-PCR primer set for human TNF or RT-PCR primer set for human ACTB (SuperArray Bioscience Corporation, Frederick, MD), SYBR Green PCR Master Mix (Applied Biosystems, Foster City, CA) and first strand DNA template.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Isolation, Real-time Polymerase Chain Reaction, Blocking Assay