Journal: NAR Cancer

Article Title: The AsiDNA™ decoy mimicking DSBs protects the normal tissue from radiation toxicity through a DNA-PK/p53/p21-dependent G1/S arrest

doi: 10.1093/narcan/zcae011

Figure Lengend Snippet: AsiDNA™ induces a G1/S arrest in healthy cells in vitro . Cells were pulse-labelled with BrdU following incubation with 20 and 40 μM of AsiDNA™ for 24 and 48 h. Representative images of the bivariate analysis by flow cytometry of BrdU incorporation versus DNA content (PI) in ( A ) BJ and ( B ) RPE-hTERT cells. The deconvolution of the cellular DNA content frequency histograms allows the identification of G1 phase (purple), S-phase (orange), and G2/M-phase (green) in ( C ) BJ, and ( D ) RPE-hTERT cells. The percentage of cells in G1, S, and G2/M is shown in ( E ) for BJ, and in ( F ) for RPE-hTERT cells. Data are expressed as mean ± standard deviation ( n = 8–9) with significance given by two-way ANOVA, Tukey's multiple comparison tests, and represented above the bar plots. Statistical significance was set at * P value < 0.05, ** P value < 0.01, *** P value < 0.001 and **** P value < 0.0001.

Article Snippet: Immortalized retinal pigment epithelial cell line hTERT (RPE-hTERT, kindly provided by A. Londono, Institut Curie, France), RPE-hTERT with shp53 (kindly provided by D. Fachinetti, Institut Curie, France), immortalized primary fibroblasts hTERT (VH10-hTERT, kindly provided by Aart G Jochemsen, and described in ( )), immortalized RPE-hTERT p21 −/− (kindly provided by R. G. Syljuåsen and described in ( )), primary human skin fibroblasts (BJ, ATCC CRL-2522), primary human lung fibroblasts (MRC-5, kindly provided by P. Jeggo, GDSC, Brighton, UK), and SV40-transformed MRC-5 fibroblasts (MRC-5v1, kindly provided by P. Jeggo, GDSC, Brighton, UK) were cultured in DMEM/F12 glutamaxTM supplement medium (Thermo Fisher Scientific, France) supplemented with 10% fetal calf serum (FCS, Eurobio, France) and 100U/ml penicillin 100 μg/ml streptomycin (P/S, Thermo Fisher Scientific, France).

Techniques: In Vitro, Incubation, Flow Cytometry, BrdU Incorporation Assay, Standard Deviation, Comparison

Journal: NAR Cancer

Article Title: The AsiDNA™ decoy mimicking DSBs protects the normal tissue from radiation toxicity through a DNA-PK/p53/p21-dependent G1/S arrest

doi: 10.1093/narcan/zcae011

Figure Lengend Snippet: AsiDNA™-induced G1/S arrest in vitro is dependent on DNA-PK, p53, and p21. ( A ) RPE-hTERT cells were pre-treated with 1 μM olaparib (Ola) or 10 μM NU7026 (NU) for 1 h before addition of 20 μM AsiDNA™. The percentage of cells in G1, S and G2/M was analysed by flow cytometry 48 h post-AsiDNA™ treatment based on PI staining. ( B ) RPE-hTERT cells were transiently transfected with small inhibitory RNA (siRNA) silencing DNA-PKcs, p53, or p21 before being exposed to AsiDNA™ for 48 h. The percentage of cells in G1, S and G2/M was analysed by flow cytometry based on PI staining. ( C ) MRC-5V1, ( D ) RPE-hTERT shp53 and ( E ) RPE-hTERT p21 −/− cells were treated with 20 and 40 μM of AsiDNA™ for 24 and 48 h. The percentage of cells in G1, S and G2/M was analysed by flow cytometry at the end of AsiDNA™ treatment based on PI staining. All the data are expressed as mean ± standard deviation ( n = 3–9) with significance given by two-way ANOVA, Tukey's multiple comparison tests, and represented above the bar plots. Statistical significance was set at * P value < 0.05, ** P value < 0.01, *** P value < 0.001 and **** P value < 0.0001.

Article Snippet: Immortalized retinal pigment epithelial cell line hTERT (RPE-hTERT, kindly provided by A. Londono, Institut Curie, France), RPE-hTERT with shp53 (kindly provided by D. Fachinetti, Institut Curie, France), immortalized primary fibroblasts hTERT (VH10-hTERT, kindly provided by Aart G Jochemsen, and described in ( )), immortalized RPE-hTERT p21 −/− (kindly provided by R. G. Syljuåsen and described in ( )), primary human skin fibroblasts (BJ, ATCC CRL-2522), primary human lung fibroblasts (MRC-5, kindly provided by P. Jeggo, GDSC, Brighton, UK), and SV40-transformed MRC-5 fibroblasts (MRC-5v1, kindly provided by P. Jeggo, GDSC, Brighton, UK) were cultured in DMEM/F12 glutamaxTM supplement medium (Thermo Fisher Scientific, France) supplemented with 10% fetal calf serum (FCS, Eurobio, France) and 100U/ml penicillin 100 μg/ml streptomycin (P/S, Thermo Fisher Scientific, France).

Techniques: In Vitro, Flow Cytometry, Staining, Transfection, Standard Deviation, Comparison

Journal: NAR Cancer

Article Title: The AsiDNA™ decoy mimicking DSBs protects the normal tissue from radiation toxicity through a DNA-PK/p53/p21-dependent G1/S arrest

doi: 10.1093/narcan/zcae011

Figure Lengend Snippet: AsiDNA™ treatment protects p53-proficient normal cells, but not p53-proficient tumour cells, from radiation-induced toxicity in vitro . ( A ) p53 proficient normal cells (BJ and RPE-hTERT), ( B ) p53 deficient normal cells (RPE-hTERT shp53) and ( C ) p53 proficient tumour (HCT116 and A549) cells were pre-treated with AsiDNA™ 24 h before being co-exposed to increased doses of ionizing radiation (0–6 Gy). The survival fraction was determined 8–12 days post-treatment using a clonogenic survival assay. Data are expressed as mean ± standard deviation ( n = 3 for BJ, RPE-hTERT shp53, HCT116 and A549; n = 4 for RPE-hTERT), fitted to the linear-quadratic model as a function of dose with significance given by nonlinear fit using GraphPad Prism.

Article Snippet: Immortalized retinal pigment epithelial cell line hTERT (RPE-hTERT, kindly provided by A. Londono, Institut Curie, France), RPE-hTERT with shp53 (kindly provided by D. Fachinetti, Institut Curie, France), immortalized primary fibroblasts hTERT (VH10-hTERT, kindly provided by Aart G Jochemsen, and described in ( )), immortalized RPE-hTERT p21 −/− (kindly provided by R. G. Syljuåsen and described in ( )), primary human skin fibroblasts (BJ, ATCC CRL-2522), primary human lung fibroblasts (MRC-5, kindly provided by P. Jeggo, GDSC, Brighton, UK), and SV40-transformed MRC-5 fibroblasts (MRC-5v1, kindly provided by P. Jeggo, GDSC, Brighton, UK) were cultured in DMEM/F12 glutamaxTM supplement medium (Thermo Fisher Scientific, France) supplemented with 10% fetal calf serum (FCS, Eurobio, France) and 100U/ml penicillin 100 μg/ml streptomycin (P/S, Thermo Fisher Scientific, France).

Techniques: In Vitro, Clonogenic Cell Survival Assay, Standard Deviation

Journal: Marine Drugs

Article Title: 4-(Phenylsulfanyl) Butan-2-One Attenuates the Inflammatory Response Induced by Amyloid-β Oligomers in Retinal Pigment Epithelium Cells

doi: 10.3390/md19010001

Figure Lengend Snippet: The effects of oAβ 1-42 and 4-PSB-2 on the viability of human adult retinal pigment epithelial cell line (ARPE-19 cells). ( A ) The expression of oAβ 1-42 in ARPE-19 cells: A11 (red), oAβ marker; DAPI (blue), nucleus; Bar, 100 µm. ( B ) A dose–response curve for RPE cells stimulated with 0.1 to 10 µM solutions of oAβ 1-42 for 48 h demonstrates the mild effect of 10 µM oAβ 1-42 on the viability of ARPE-19 cells ( n = 5–6/group). ( C ) The morphology of ARPE-19 cells after 0.1 to 10 µM of oAβ 1-42 treatment compared to control; Bar, 20 µm. The irregular shapes of cell bodies and nuclei of ARPE-19 cells (black arrows) and small vesicles in the cytoplasm (white arrows) were observed after oAβ 1-42 treatment. ( D ) Chemical structure of 4-PSB-2. ( E ) ARPE-19 cells were treated with 0.1% dimethyl sulfoxide (DMSO) in control, 4-PSB-2 (1, 25, 50, 100, and 200 µM) for 24 h. MTT analysis showed that 4-PSB-2 was not toxic to ARPE-19 cells, and 25 µM of 4-PSB-2 significantly increased cell viability.(* indicates p ≤ 0.001 between the groups).

Article Snippet: A human retinal pigment epithelial cell line (ARPE-19) was provided by Rong-Kung Tsai at Institute of Medical Sciences, Tzu Chi University (Taiwan), and cultured in Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (DMEM/F12), containing 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in 5% CO 2 and 95% air.

Techniques: Expressing, Marker, Control