Journal: International Journal of Molecular Sciences

Article Title: Tempol Exerts Radioprotective Effects by Suppressing Radiation-Induced DNA Double-Strand Break Formation

doi: 10.3390/ijms27062601

Figure Lengend Snippet: Radiation-induced ROS production and DSB formation are suppressed in tempol-treated cells. HeLa or TIG-3 cells were cultured in medium containing 50 µmol/L tempol for 1 week and irradiated with 1 Gy γ-rays or particle beams. ( A ) Intracellular ROS levels in HeLa cells until 6 h after 1 Gy radiation. ( B ) Average number of γ-H2AX foci per cell in HeLa cells after 1 Gy of γ-ray or C-ion exposure. Error bars indicate standard deviation. (γ-ray: n ≥ 100, C-ion: n ≥ 50) * p < 0.05 ** p < 0.01. ( C ) Average number of γ-H2AX foci in TIG-3 cells after X-ray and Fe-ion irradiation. Error bars indicate standard deviation. (X-ray: n ≥ 100 Fe-ion: n ≥ 50) ** p < 0.01.

Article Snippet: They were then fixed with 2% paraformaldehyde for 20 min and immunostained with rabbit polyclonal anti-γ-H2AX antibody (1:500; NB100-368, Novus, Centennial, CO, USA), rat monoclonal anti-F4/80 antibody (1:500; ab6640, abcam, Cambridge, UK) and rabbit polyclonal anti-Annexin V antibody (1:200; ab14196, abcam, Cambridge, UK), respectively.

Techniques: Cell Culture, Irradiation, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Tempol Exerts Radioprotective Effects by Suppressing Radiation-Induced DNA Double-Strand Break Formation

doi: 10.3390/ijms27062601

Figure Lengend Snippet: Tempol suppresses radiation-induced DSB formation in mouse tissues. Mice were fed either a normal diet or a diet containing Tempol (10 mg/g) for 5 d, followed by exposure to 1 Gy γ-rays. The thymus and duodenum of the mice were collected at the following time points: immediately after irradiation (0 h), 1 h and 6 h. ( A ) Fluorescence intensity of γ-H2AX in mouse thymus after 1 Gy γ-ray irradiation. Error bars indicate mean ± SEM from two independent experiments using a total of 3–4 mice (1Gy-1hr: n = 4, others: n = 3). arb.u.: arbitrary unit. ( B ) Average number of γ-H2AX foci per cell in mouse duodenum after 1 Gy γ-ray irradiation. Error bars indicate standard deviation. (1Gy-1hr: n = 4, others: n = 3). p -value indicates significant differences compared with Tempol (−).

Article Snippet: They were then fixed with 2% paraformaldehyde for 20 min and immunostained with rabbit polyclonal anti-γ-H2AX antibody (1:500; NB100-368, Novus, Centennial, CO, USA), rat monoclonal anti-F4/80 antibody (1:500; ab6640, abcam, Cambridge, UK) and rabbit polyclonal anti-Annexin V antibody (1:200; ab14196, abcam, Cambridge, UK), respectively.

Techniques: Irradiation, Fluorescence, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Tempol Exerts Radioprotective Effects by Suppressing Radiation-Induced DNA Double-Strand Break Formation

doi: 10.3390/ijms27062601

Figure Lengend Snippet: Tempol suppresses DSB formation after chronic irradiation. ( A ) HeLa cells were cultured in medium containing 50 µmol/L tempol for 1 week, followed by fixing and staining for γ-H2AX at 0, 1, 2, and 6 h after 1 Gy γ-ray irradiation. The graph shows the mean number of γ-H2AX foci. Error bars indicate standard deviation ( n ≥ 100) * p < 0.05, ** p < 0.01. ( B , C ) Mice were fed either a normal diet or a diet containing tempol and exposed to 1 Gy γ-rays (0.694 mGy/min) for 24 h. Tissue samples were obtained from mice at 0 h, 1 h, and 6 h. The graph shows the fluorescence intensity of γ-H2AX in the mouse thymus B. and the average number of γ-H2AX foci in the mouse duodenum after chronic γ-ray irradiation C. The error bars indicate mean ± SEM from two independent experiments ( B ) or the standard deviation ( C ). (1Gy-1hr: n = 4, others: n = 3) p- value indicates significant differences compared with Tempol (−). * p < 0.05. arb.u.: arbitrary unit.

Article Snippet: They were then fixed with 2% paraformaldehyde for 20 min and immunostained with rabbit polyclonal anti-γ-H2AX antibody (1:500; NB100-368, Novus, Centennial, CO, USA), rat monoclonal anti-F4/80 antibody (1:500; ab6640, abcam, Cambridge, UK) and rabbit polyclonal anti-Annexin V antibody (1:200; ab14196, abcam, Cambridge, UK), respectively.

Techniques: Irradiation, Cell Culture, Staining, Standard Deviation, Fluorescence

Journal: Nature communications

Article Title: UPF1 plays critical roles in early B cell development.

doi: 10.1038/s41467-024-50032-6

Figure Lengend Snippet: Fig. 4 | UPF1 is necessary for Igh VH-DHJH recombination at early LPre-B stage. a–d The transcript amount of Ighv (a), Ighm (b), Ighj (c), and Ighd (d) in Upf1-cKO and Ctrl early LPre-B cells. The data were obtained from the RNA-seq shown in Fig. 2 (n = 3). Each bar represents the mean ± SD from biological replicates. e Schematic illustration of germline Igh locus. Colored arrows indicate each primer used in PCR analysis in (f). f PCR assay amplifying recombined Igh as shown in (e) using the DNA derived from Upf1-cKO and Ctrl early LPre-B cells. Genome DNA derived from the tail of the WT mouse was used as a negative control. Results are representative of at

Article Snippet: The following primary antibodies were used for immunoblot analysis: antibody to UPF1 (NBP1-05967, Novus biologicals), Phospho-UPF1(071016, Merck), β-actin (sc-1615, Santa Cruz) The complete list of antibodies used in this study can be found in Supplementary table 2.

Techniques: RNA Sequencing, Derivative Assay, Negative Control

Journal: Journal of Cancer

Article Title: Amlexanox and UPF1 Modulate Wnt Signaling and Apoptosis in HCT-116 Colorectal Cancer Cells

doi: 10.7150/jca.28331

Figure Lengend Snippet: Upregulation of UPF1 in HCT-116 cells. (A) Representative Western blot showing expression of UPF1, CBP, and p300. (B) NMD assayed by the reporter system as in Fig. . Data are from nine independent experiments. (C) Wnt signaling assayed as in Fig. . (D) The fold up-regulation of Wnt activity after exposure to butyrate from the data of (C). (E) Apoptosis assays were performed as previously described - . For (C-E), data from three independent experiments are shown. Bars, SDs

Article Snippet: Control and Rent1 (UPF1) CRISPR activation vectors were from Santa Cruz Biotechnology.

Techniques: Western Blot, Expressing, Activity Assay