mouse anti 53bp1  (Novus Biologicals)

Journal: International Journal of Nanomedicine

Article Title: Antioxidant Nanoparticles Restore Cisplatin-Induced Male Fertility Defects by Promoting MDC1-53bp1-Associated Non-Homologous DNA Repair Mechanism and Sperm Intracellular Calcium Influx

doi: 10.2147/IJN.S408623

Figure Lengend Snippet: Antioxidant nanoparticle promoted MDC1-53bp1 associated non-homologous DNA break repair. ( A ) GO analysis showed that in total, 420 genes that were known to involve in the signaling pathways related to wound response, tissue remodeling, wound healing, and apoptosis regulation were highly affected. ( B ) DNA double-strand break checkpoint protein MDC1 was highly expressed in the control nHNK and nHNK treatment groups, indicating that the testicular DNA damage checkpoint and repair mechanism was functional in these groups. In contrast, this mechanism was abolished in the cisplatin group. ( C ) Quantitative data supported the immunofluorescent observation that a significant decrease in the % of MDC1-positive cells (3.5%) was measured in the cisplatin group. And nHNK treatment reversed the protein expression of MDC1 in the testis (34.6%). ( D ) 53bp1-mediated non-homologous DNA repair mechanism was activated upon nHNK treatment, and no changes can be detected regarding the BRAC1-associated homologous DNA repair mechanism. Bars represent standard deviation (S.D.), and results were presented as mean ± standard deviation (S.D.). *p<0.05, ***p<0.001, ****p<0.0001.

Article Snippet: Mouse monoclonal anti-53bp1 antibody (#NBP2-25028) and anti-BRAC1 antibody (#NB100-404) were obtained from Novus Biologicals, LLC (Centennial, CO, USA).

Techniques: Protein-Protein interactions, Control, Functional Assay, Expressing, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Antioxidant Nanoparticles Restore Cisplatin-Induced Male Fertility Defects by Promoting MDC1-53bp1-Associated Non-Homologous DNA Repair Mechanism and Sperm Intracellular Calcium Influx

doi: 10.2147/IJN.S408623

Figure Lengend Snippet: Summarized findings of the current study. Cisplatin results in the overproduction of free radicals and leads to oxidative stress and damage in the testis. Accumulation of excessive free radicals leads to DNA double-strand breaks that disrupt normal spermatogenesis. Cisplatin-induced oxidative stress also causes structural and functional defects of sperm mitochondria, which further compromise normal ATP production. In combination with insufficient ATP and defected calcium channel, the calcium influx required for sperm hypermotility was likely affected as sperm progressive motility was decreased significantly, and sperm motility was affected. Natural polyphenol extracts honokiol encapsulated within nanoparticles serve as a ROS scavenger that overcomes mitochondrial oxidative damage and promotes 53bp1-associated non-homologous DNA repair mechanism. With a yet-uncovered mechanism promoting sperm calcium influx, nHNK treatment mitigates cisplatin-induced male fertility defects.

Article Snippet: Mouse monoclonal anti-53bp1 antibody (#NBP2-25028) and anti-BRAC1 antibody (#NB100-404) were obtained from Novus Biologicals, LLC (Centennial, CO, USA).

Techniques: Functional Assay

Journal: Cells

Article Title: CHRDL1 Regulates Stemness in Glioma Stem-like Cells

doi: 10.3390/cells11233917

Figure Lengend Snippet: CHRDL1-depletion radiosensitizes GSCs. ( A ) 53BP1 foci assay 24 h after irradiation of NCH644 shCtrl or NCH644 shCHRDL1 GSCs with the indicated doses. ( B ) Sphere formation of NCH644 shCtrl or shCHRDL1 cells 7 days after seeding of single cells and irradiation as indicated. ( C ) Representative microphotographs of NCH644 shCtrl or NCH644 shCRHDL1 without irradiation (0 Gy) or after irradiation with 6 Gy; scale bar: 200 µm. * p < 0.05; ** p < 0.01; **** p < 0.0001; two-way-ANOVA with Sidak’s multiple comparison tests. ( D ) Sphere formation of NCH644 treated with solvent or recombinant human BMP4 (25 ng/mL) immediately prior to irradiation with a dose of 4 or 6 Gy. Sphere area was determined 7 days after treatment. * p < 0.05; **** p < 0.0001; Kruskal—Wallis test with Dunn’s multiple comparisons test.

Article Snippet: The following primary and secondary antibodies were used: 53BP1 (NP-100-304, Bio-Techne GmbH, Wiesbaden, Germany), 1:1000; Alexa Fluor ® 488 F(ab’)2 fragment goat anti-rabbit IgG (H + L) (Thermo Fisher); 1:500.

Techniques: Irradiation, Comparison, Solvent, Recombinant