Journal: Journal of Radiation Research

Article Title: Impact of time interval and dose rate on cell survival following low-dose fractionated exposures

doi: 10.1093/jrr/rrx025

Figure Lengend Snippet: Relative survivals of V79 (a) and A549 (b) cells after fractionated exposures with varying dose rates. Total doses of 2 Gy were administered by fractionated radiation in unit doses of 0.25, 0.5, 1.0 and 2.0 Gy at 1.0, 1.5 and 2.0 Gy/min with 1 min intervals (Irradiation Method 1). * P < 0.05 vs 2 Gy × 1 (each condition), ** P < 0.01 vs 2 Gy × 1 (each condition). A single dagger ( † ) indicates P < 0.05. Double daggers ( †† ) indicate P < 0.05. Data represent the mean ± standard deviation of the results of three to five independent experiments, each with four samples.

Article Snippet: Cells were cultured in DMEM/Ham's F-12 (A549) and Ham's F-12 (V79) media (Wako, Japan) supplemented with 10% fetal bovine serum, and were maintained at 37°C with 95% air and 5% CO 2 .

Techniques: Irradiation, Standard Deviation

Journal: Journal of Radiation Research

Article Title: Impact of time interval and dose rate on cell survival following low-dose fractionated exposures

doi: 10.1093/jrr/rrx025

Figure Lengend Snippet: Relative survivals of V79 (a) and A549 (b) cells after fractionated exposures with varying time intervals. Total doses of 2 Gy were administered by fractionated radiation in unit doses of 0.25, 0.5, 1.0 and 2.0 Gy at 2.0 Gy/min with intervals of 10 s, 1 min and 3 min (Irradiation Method 2). * P < 0.05 vs 2 Gy × 1 (each condition), ** P < 0.01 vs 2 Gy × 1 (each condition). A single dagger ( † ) indicates P < 0.05. Double daggers ( †† ) indicate P < 0.05. Data represent the mean ± standard deviation of the results of three to five independent experiments, each with four samples.

Article Snippet: Cells were cultured in DMEM/Ham's F-12 (A549) and Ham's F-12 (V79) media (Wako, Japan) supplemented with 10% fetal bovine serum, and were maintained at 37°C with 95% air and 5% CO 2 .

Techniques: Irradiation, Standard Deviation

Journal: Journal of Radiation Research

Article Title: Impact of time interval and dose rate on cell survival following low-dose fractionated exposures

doi: 10.1093/jrr/rrx025

Figure Lengend Snippet: Relative survivals of V79 (a) and A549 (b) cells after fractionated exposures with varying total doses. Total doses of 2, 4 and 8 Gy were administered by fractionated radiation in unit doses of 0.25, 0.5, 1.0 and 2.0 Gy at 2.0 Gy/min with 10-s intervals (Irradiation Method 3). * P < 0.05 vs Total dose (each condition), ** P < 0.01 vs Total dose (each condition). Data represent the mean ± standard deviation of the results of three to five independent experiments, each with four samples.

Article Snippet: Cells were cultured in DMEM/Ham's F-12 (A549) and Ham's F-12 (V79) media (Wako, Japan) supplemented with 10% fetal bovine serum, and were maintained at 37°C with 95% air and 5% CO 2 .

Techniques: Irradiation, Standard Deviation

Journal: Journal of Radiation Research

Article Title: Impact of time interval and dose rate on cell survival following low-dose fractionated exposures

doi: 10.1093/jrr/rrx025

Figure Lengend Snippet: Cell survival curves of V79 (a) and A549 (b) cells after fractionated exposure with varying unit doses. Total doses of 1, 2, 4, 6 and 8 Gy were administered by fractionated irradiation in unit doses of 0.25, 0.5, 1.0 and 2.0 Gy at 2.0 Gy/min with 10-s intervals. Curves were fitted to the linear–quadratic model. Data represent the mean ± standard deviation of the results of three to five independent experiments, each with four samples.

Article Snippet: Cells were cultured in DMEM/Ham's F-12 (A549) and Ham's F-12 (V79) media (Wako, Japan) supplemented with 10% fetal bovine serum, and were maintained at 37°C with 95% air and 5% CO 2 .

Techniques: Irradiation, Standard Deviation

Journal: Journal of Radiation Research

Article Title: Impact of time interval and dose rate on cell survival following low-dose fractionated exposures

doi: 10.1093/jrr/rrx025

Figure Lengend Snippet: Values of the parameters obtained from survival curves of V79 and A549 cells using the linear-quadratic model

Article Snippet: Cells were cultured in DMEM/Ham's F-12 (A549) and Ham's F-12 (V79) media (Wako, Japan) supplemented with 10% fetal bovine serum, and were maintained at 37°C with 95% air and 5% CO 2 .

Techniques:

Journal: Journal of Radiation Research

Article Title: Impact of time interval and dose rate on cell survival following low-dose fractionated exposures

doi: 10.1093/jrr/rrx025

Figure Lengend Snippet: Apoptosis was analyzed by flow cytometry using Annexin V and PI in V79 (a) and A549 (b) cells at 48 h after irradiation. No significant differences in apoptosis were observed between low-dose and single fractionated exposures in either V79 or A549 cells. Data represent the mean ± standard deviation of the results of three independent experiments.

Article Snippet: Cells were cultured in DMEM/Ham's F-12 (A549) and Ham's F-12 (V79) media (Wako, Japan) supplemented with 10% fetal bovine serum, and were maintained at 37°C with 95% air and 5% CO 2 .

Techniques: Flow Cytometry, Irradiation, Standard Deviation

Journal: Journal of Radiation Research

Article Title: Impact of time interval and dose rate on cell survival following low-dose fractionated exposures

doi: 10.1093/jrr/rrx025

Figure Lengend Snippet: Cell cycle analysis of V79 (a, c) and A549 (b, d) cells by flow cytometer using PI at 12 and 24 h after irradiation. Each value in the distribution is the mean of four to eight independent experiments. (e) A representative flow cytometric histogram of PI at 12 and 24 h after irradiation. The cell cycle distribution was not significantly altered between low dose and single fractionated exposures in either V79 or A549 cells.

Article Snippet: Cells were cultured in DMEM/Ham's F-12 (A549) and Ham's F-12 (V79) media (Wako, Japan) supplemented with 10% fetal bovine serum, and were maintained at 37°C with 95% air and 5% CO 2 .

Techniques: Cell Cycle Assay, Flow Cytometry, Irradiation

Journal: The Journal of Biological Chemistry

Article Title: Mechanism of Folding and Activation of Subtilisin Kexin Isozyme-1 (SKI-1)/Site-1 Protease (S1P) *

doi: 10.1074/jbc.M115.677757

Figure Lengend Snippet: Characterization of the activity of the prodomain deletion mutants. A, SREBP-2 processing by SKI-1/S1P mutants. SRD12B cells were transfected with the indicated SKI-1/S1P constructs. After 32 h, the SREBP-2 pathway was induced by mevastatin treatment for 18 h, whereas control cells were treated with DMSO. After treatment, total RNA was extracted to analyze HGMCS1 gene induction by quantitative RT-PCR. The results were analyzed by the ΔΔCT method, and the data were normalized against hydroxymethylbilane synthase gene expression. Gene expression is represented as fold induction above levels for control treatment (pIR-HA DMSO) (mean ± S.D.; n = 3). B, expression of SKI-1/S1P variants in A was assessed by Western blot as in Fig. 1B. C and D, LASV and LCMV GPC processing by the SKI-1/S1P mutants. SRD12B cell were co-transfected with either LASV GPC (C) or LCMV GPC (D) and the indicated SKI-1/S1P variants. At 48 h post-transfection, the cell lysates were analyzed by Western blot to assess GPC processing. SKI-1/S1P expression was detected with anti-V5 antibody and tubulin (Tub) detected as loading control. Tubulin, the precursor GPC, mature GP2, and maturation forms of SKI-1/S1P (forms A, B, and C) are indicated. E, schematic of the SKI-1/S1P sensor bearing the cleavage motif of LASV GPC. The signal peptide (SP), the GLuc reporter, the LASV GPC-derived cleavage motif, and the SKI-1/S1P-derived stump region are indicated. F, processing of the SKI-1/S1P sensor by SKI-1/S1P mutants. SRD12B cells were co-transfected with the SKI-1/S1P sensor (SS-LASV) and the indicated SKI-1/S1P variants. At 48 h post-transfection, cell lysates and supernatants were collected and analyzed by Western blot using an anti-GLuc antibody (bottom). Cleaved sensor (cGLuc) and the uncleaved precursor (pGLuc) are indicated. Conditioned media were analyzed for GLuc activity by addition of coelenterazine substrate. The data are shown as relative light units (RLU) (means ± S.D.; n = 3).

Article Snippet: Drug Treatments Induction of genes regulated by SREBP2 was triggered by treatment with Ham's F-12 DMEM 1:1 containing 5% (v/v) delipidated FBS, 50 μ m sodium mevalonate, and 50 μ m mevastatin (Enzo Lifescience) for 18 h, as reported previously ( 20 ).

Techniques: Activity Assay, Transfection, Construct, Quantitative RT-PCR, Expressing, Western Blot, Derivative Assay