Journal: Viruses

Article Title: Disruption of Zika Virus xrRNA1-Dependent sfRNA1 Production Results in Tissue-Specific Attenuated Viral Replication

doi: 10.3390/v12101177

Figure Lengend Snippet: In vitro sfRNA production and viral growth kinetics of ZIKV X1 compared to WT ZIKV clone. ( A ) A549 cells were infected with either X1 or WT clone at a MOI of 1. At 48 h post infection (HPI), cellular RNA was collected and the presence of ZIKV sfRNAs was detected in two biological replicates per infection via Northern blot using a ZIKV 3′ UTR-specific probe. To analyze viral growth kinetics, human A549 cells ( B ), ( C ) or Aedes albopictus U4.4 cells ( D ), ( E ) were infected with X1 or WT at an MOI of 0.1. At 0, 24, 48, and 72 HPI, supernatant was collected and used to measure either extracellular viral RNA via RT-qPCR ( B – D ) or infectious virus via FFU assay ( C – E ). ( B – E ) Dashed lines represent the limit of detection (LOD). Error bars indicate standard error of the mean for six replicates across two independent experiments. ( n = 6, NS by two-way ANOVA).

Article Snippet: African green monkey kidney epithelial cells (Vero E6), human lung epithelial cells (A549), and Aedes albopictus cells (C6/36) were sourced from the American Type Culture Collection (ATCC).

Techniques: In Vitro, Infection, Northern Blot, Quantitative RT-PCR, Virus

Journal: Investigational New Drugs

Article Title: Blood glutamate scavengers prolong the survival of rats and mice with brain-implanted gliomas

doi: 10.1007/s10637-012-9794-x

Figure Lengend Snippet: a . A representative example of MRI analysis of C6 glioma growth in vivo. At 7 days post-implantation and before the treatment, an MRI imaging was performed (left column, 1–2 pictures T2 imaging, 3–4 pictures T1 imaging). The rats were randomized and either treated with 0.2 M OxAc or 0.3 M NaCl for additional 14 days. The MRI imaging was performed again at the end of the treatment (21 days post glioma implantation, right column, 1–4 pictures T2 imaging). b . Tumor volume of the rats before and after the treatment was calculated in n = 14 for control group and in n = 15 for the treated group using MRIcro software. OxAc-treated rats show significantly reduced tumor growth compared to the control group. * p < 0.01 (repeated measures ANOVA test)

Article Snippet: Rat C6 glioma and human U-87 Glioma cells were obtained from ATCC.

Techniques: In Vivo, Imaging, Control, Software

Journal:

Article Title: Perturbation of Hyaluronan Interactions Inhibits Malignant Properties of Glioma Cells

doi:

Figure Lengend Snippet: Inhibition of anchorage-independent growth of C6 glioma cells by hyaluronan oligomers or by overexpression of soluble HABPs. A: C6 cells (2 × 103 per well) were incubated for 1 week in soft agar in the presence or absence of 1, 10, or 100 μg/ml of hyaluronan oligomers, or 50 μg/ml of N-acetylglucosamine plus 50 μg/ml of glucuronic acid monomers, as described in Materials and Methods. The numbers of colonies >0.25 mm in size were counted. The results are presented as means of triplicates ± SD (control versus 10 or 100 μg/ml hyaluronan oligomers; P < 0.05). B: Western blot with antibody against phosphorylated Akt (p-Akt) or Akt for extracts of C6 cells grown in suspension. Lane 1, C6 cells alone; lane 2, C6 cells incubated with 100 μg/ml of chitin oligomers; lane 3, with 100 μg/ml of hyaluronan polymer (∼80 kD); lane 4, with 100 μg/ml of hyaluronan oligomers. C: C6 cells (2 × 103 per well) were infected with recombinant adenoviruses driving expression of β-galactosidase (β-gal), soluble CD44 (solCD44), or brevican link module (BLM), then assayed for growth of colonies in soft agar. The results are presented as means of three separate experiments, each performed in triplicate, ± SD (control versus soluble CD44 or brevican link module; P < 0.05).

Article Snippet: C6 rat glioma cells, A172 human glioma cells, and U87 human glioma cells were obtained from the American Type Culture Collection (Rockville, MD).

Techniques: Inhibition, Over Expression, Incubation, Control, Western Blot, Suspension, Polymer, Infection, Recombinant, Expressing

Journal:

Article Title: Perturbation of Hyaluronan Interactions Inhibits Malignant Properties of Glioma Cells

doi:

Figure Lengend Snippet: Inhibition of glioma cell invasion by hyaluronan oligomers. A: C6 glioma cells (2 × 104 per well) were incubated in the presence or absence of 100 μg/ml of hyaluronan (HA) oligomers for 72 hours and the number of cells that invaded the reconstituted basement membrane matrix was measured at each 24-hour interval, as described in Materials and Methods. B: C6 rat glioma cells (2 × 104 per well), A172 human glioma cells (1 × 104 per well), and U87 human glioma cells (1 × 104 per well) were incubated in the presence and absence of 1 to 100 μg/ml of hyaluronan oligomers, 100 μg/ml of chitin oligomers, or 4 μg/ml of antibody against CD44 (Zymed), and the number of cells that invaded was measured after 24 hours. The results in B are presented as means of three experiments ± SD (control versus hyaluronan oligomers; P < 0.05 for each cell type).

Article Snippet: C6 rat glioma cells, A172 human glioma cells, and U87 human glioma cells were obtained from the American Type Culture Collection (Rockville, MD).

Techniques: Inhibition, Incubation, Membrane, Control

Journal: Cell Death Discovery

Article Title: TRAIL DR5-CTSB crosstalk participates in breast cancer autophagy initiated by SAHA

doi: 10.1038/cddiscovery.2017.52

Figure Lengend Snippet: The effects of SAHA and TRAIL DR5 on the expressions of autophagy-related genes in cancer cells. RNA was isolated from MDA-MB-231 or MCF-7 cells using high pure RNA isolation kit following manufacturer’s instructions. To obtain the first-strand cDNA, transcriptor first strand cDNA synthesis kit was used and cDNA was as a template in real-time PCR reactions with power SYBR green PCR master mix. Exprofile human autophagy Gene qPCR array kit was employed to describe related mRNA expression. Relative changes of gene expression in the array were calculated using the 2 −ΔΔCt (threshold cycle) method. ( a – c ). MDA-MB-231 cells, ( d – f ). MCF-7 cells, ( g ). Related data analysis. Relative quantitative values represent mean±S.E.M.; * represents statistical significance of P <0.05 comparing with Basal, ** represents statistical significance of P <0.01 comparing with Basal.

Article Snippet: Exprofile human autophagy gene qPCR array kit was obtained from Genecopoeia (Rockville, MD.

Techniques: Isolation, cDNA Synthesis, Real-time Polymerase Chain Reaction, SYBR Green Assay, Expressing, Gene Expression

Journal: RSC Advances

Article Title: Developing a far-red fluorogenic beta-galactosidase probe for senescent cell imaging and photoablation †

doi: 10.1039/d2ra00377e

Figure Lengend Snippet: Cellular characterization in C6 (β-gal-negative) and C6/ LacZ (β-gal-positive) cells. (A) X-gal staining to determine β-gal expression (magnification: ×20). (B) Cellular uptake and activation of Gal-MB. Both cells were stained with Hoechst 33342 after treating with Gal-MB (5 μM) for 1 h (magnification: ×40).

Article Snippet: C6 (rat glioma), C6/ LacZ ( LacZ transfected C6 cell line), MDA-MB231 human breast cancer cell line, and A549 human non-small-cell lung carcinoma cell line were purchased from American Type Culture Collection (ATCC; Manassas, VA) and maintained in Dulbecco's modified Eagle's medium (DMEM) medium (Mediatech Inc., Manassas, VA) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich), and 1% penicillin/streptomycin in an incubator with humidified air containing 5% CO 2 at 37 °C.

Techniques: Staining, Expressing, Activation Assay

Journal: RSC Advances

Article Title: Developing a far-red fluorogenic beta-galactosidase probe for senescent cell imaging and photoablation †

doi: 10.1039/d2ra00377e

Figure Lengend Snippet: Photodynamic effect of Gal-MB in β-gal expressing cells. (A) Cell viability of Gal-MB treated cells without or with light illumination. C6 or C6/ LacZ cells were incubated with various concentration of Gal-MB (0–30 μM) for 2 h, washed, and illuminated without or with 665 nm LED light (30 mW cm −2 ) for 30 min. Cell viability assay was performed after 1 day. (B) Selective ablation of β-gal expressing LacZ cells. C6 and C6/ LacZ cells were cocultured at 7 : 3 ratio. Cells were treated with Gal-MB (0, 30, 50 μM) for 2 h and illuminated without or with LED light (30 min, 30 mW cm −2 ). Cells were cultured for additional 24 h before X-gal staining (magnification: ×20).

Article Snippet: C6 (rat glioma), C6/ LacZ ( LacZ transfected C6 cell line), MDA-MB231 human breast cancer cell line, and A549 human non-small-cell lung carcinoma cell line were purchased from American Type Culture Collection (ATCC; Manassas, VA) and maintained in Dulbecco's modified Eagle's medium (DMEM) medium (Mediatech Inc., Manassas, VA) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich), and 1% penicillin/streptomycin in an incubator with humidified air containing 5% CO 2 at 37 °C.

Techniques: Expressing, Incubation, Concentration Assay, Viability Assay, Cell Culture, Staining