Journal: Discover oncology

Article Title: Olaparib increases chemosensitivity by upregulating miR-125a-3p in ovarian cancer cells.

doi: 10.1007/s12672-025-02048-7

Figure Lengend Snippet: Fig. 4 Effects of miR-125a-3p overexpression induces cellular senescence and increases DNA damage. A The SPiDER-βGal staining results were detected by flow cytometry. Data represent the mean ± standard deviation (n = 3). B The effect of different expression induced DNA damage was detected by γ-H2AX staining (×200). C Using western blotting to detect the expression of the γ-H2AX protein

Article Snippet: The DNA damage (γ-H2AX) detection kit-green (G265) was purchased from Dojindo Laboratories, Inc.

Techniques: Over Expression, Staining, Flow Cytometry, Standard Deviation, Expressing, Western Blot

Journal: Discover oncology

Article Title: Olaparib increases chemosensitivity by upregulating miR-125a-3p in ovarian cancer cells.

doi: 10.1007/s12672-025-02048-7

Figure Lengend Snippet: Fig. 6 Proposed model summarizing olaparib increases chemosensitivity by upregulating miR-125a-3p. Upregulating the expression of miR-125a-3p, leading to cell senescence and G1 phase arrest of the cell cycle, then induces DNA damage and improving the therapeutic sensitivity of ovarian cancer cells

Article Snippet: The DNA damage (γ-H2AX) detection kit-green (G265) was purchased from Dojindo Laboratories, Inc.

Techniques: Expressing

Journal: Clinical and Experimental Reproductive Medicine

Article Title: The role of nGPx4 in resisting DEHP-induced DNA damage and reducing caspase‐independent cell death in male germ cells

doi: 10.5653/cerm.2024.07521

Figure Lengend Snippet: Effects of nuclear glutathione peroxidase 4 (nGPx4) overexpression on DNA damage and apoptosis-related protein expression in germ cell line (GC-1) spg cells after mono-2-ethylhexyl phthalate (MEHP) exposure. Quantitative analysis of mRNA expression levels of (A) phosphorylated H2A histone variant (γ-H2A.X), (B) Caspase-3, (C) Bcl 2-associated X (Bax), and (D) B-cell lymphoma 2 (Bcl-2), demonstrating MEHP’s impact on these genes. (E) Western blot analysis of apoptosis-related proteins. (F) Quantitative immunofluorescence analysis of the DNA damage marker γ-H2A.X. Image analysis was performed using ImageJ, and the data are presented as mean±standard deviation. Scale bar: 20 µm. ns, not significant; NC, negative control; DMSO, dimethyl sulfoxide; DAPI, 4′,6-diamidino-2-phenylindole. a) p <0.05; b) p <0.01; c) p <0.001; d) p <0.0001 compared with the control group.

Article Snippet: GC-1 cells and mouse testicular tissues were digested and lysed in radioimmunoprecipitation assay (RIPA) buffer (AR0105; Boster) containing protease inhibitors (AR1182; Servicebio) for 30 minutes to extract proteins, including AIF, γ-H2A.X, caspase-3, Bax, and B-cell lymphoma 2 (Bcl-2). nGPx4 and truncated AIF (tAIF) were isolated using a cytoplasmic and nuclear separation kit (AR0106; Boster).

Techniques: Over Expression, Expressing, Variant Assay, Western Blot, Immunofluorescence, Marker, Standard Deviation, Negative Control, Control

Journal: Clinical and Experimental Reproductive Medicine

Article Title: The role of nGPx4 in resisting DEHP-induced DNA damage and reducing caspase‐independent cell death in male germ cells

doi: 10.5653/cerm.2024.07521

Figure Lengend Snippet: Impact of nuclear glutathione peroxidase 4 (nGPx4) overexpression on truncated apoptosis-inducing factor (tAIF)/phosphorylated H2A histone variant (γ-H2A.X) axis activation and apoptosis in germ cell line (GC-1) spg cells following mono-2-ethylhexyl phthalate (MEHP) exposure. (A) Western blot analysis of AIF and γ-H2A.X protein expression. (B) Quantitative immunofluorescence analysis of the DNA damage marker AIF. (C) Expression of nGPx4. (D) Translocation of AIF from the cytoplasm to the nucleus. (E) Immunoprecipitation analysis from MEHP-treated and untreated groups using anti-γ-H2A.X/anti-immunoglobulin G (IgG) antibodies. (F) Quantitative mRNA expression analysis of nGPx4, AIF, and γ-H2A.X. (G, H) Flow cytometry analysis of apoptotic GC-1 cells following MEHP exposure. Data were analyzed using ImageJ and are expressed as mean±standard deviation. Scale bar: 20 µm. OE, overexpress; NC, negative control; IP, immunoprecipitation; IB, immunoblotting; DMSO, dimethyl sulfoxide; DAPI, 4′,6-diamidino-2-phenylindole; ns, not significant; 7-AAD, 7-aminoactinomycin D; APC, allophycocyanin-conjugated. a) p <0.01; b) p <0.001; c) p <0.0001 compared with the control group.

Article Snippet: GC-1 cells and mouse testicular tissues were digested and lysed in radioimmunoprecipitation assay (RIPA) buffer (AR0105; Boster) containing protease inhibitors (AR1182; Servicebio) for 30 minutes to extract proteins, including AIF, γ-H2A.X, caspase-3, Bax, and B-cell lymphoma 2 (Bcl-2). nGPx4 and truncated AIF (tAIF) were isolated using a cytoplasmic and nuclear separation kit (AR0106; Boster).

Techniques: Over Expression, Variant Assay, Activation Assay, Western Blot, Expressing, Immunofluorescence, Marker, Translocation Assay, Immunoprecipitation, Flow Cytometry, Standard Deviation, Negative Control, Control

Journal: Clinical and Experimental Reproductive Medicine

Article Title: The role of nGPx4 in resisting DEHP-induced DNA damage and reducing caspase‐independent cell death in male germ cells

doi: 10.5653/cerm.2024.07521

Figure Lengend Snippet: Flow cytometry analysis of acridine orange (AO) and chromomycin A3 (CMA3) data. (A) Western blot analysis of phosphorylated H2A histone variant (γ-H2A.X). (B, C) Immunofluorescence staining of γ-H2A.X in testis sections. (D) Quantitative mRNA expression analysis of γ-H2A.X. (E, F) AO orange fluorescence intensity. (G, H) CMA3 binding positivity rates. (I) Sperm chromatin status assessed by sperm chromatin structure analysis, showing DNA fragmentation index (DFI) and high DNA staining (HDS) percentages. Data are expressed as mean±standard deviation. Scale bar: 50 µm. ns, not significant; WT, wild-type; DEHP, di(2-ethyl-hexyl) phthalate; nGPx4, nuclear glutathione peroxidase 4; NC, negative control; DAPI, 4′,6-diamidino-2-phenylindole; PerCP-A, phycoerythrincy5.5; PE-A, phycoerythrin; FITC, fluorescein isothiocyanate. a) p <0.05; b) p <0.01; c) p <0.001; d) p <0.0001 compared with the control group.

Article Snippet: GC-1 cells and mouse testicular tissues were digested and lysed in radioimmunoprecipitation assay (RIPA) buffer (AR0105; Boster) containing protease inhibitors (AR1182; Servicebio) for 30 minutes to extract proteins, including AIF, γ-H2A.X, caspase-3, Bax, and B-cell lymphoma 2 (Bcl-2). nGPx4 and truncated AIF (tAIF) were isolated using a cytoplasmic and nuclear separation kit (AR0106; Boster).

Techniques: Flow Cytometry, Western Blot, Variant Assay, Immunofluorescence, Staining, Expressing, Fluorescence, Binding Assay, Standard Deviation, Negative Control, Control

Journal: Clinical and Experimental Reproductive Medicine

Article Title: The role of nGPx4 in resisting DEHP-induced DNA damage and reducing caspase‐independent cell death in male germ cells

doi: 10.5653/cerm.2024.07521

Figure Lengend Snippet: Di (2-ethyl-hexyl) phthalate (DEHP) exposure induces caspase-independent cell death in male mouse germ cells via the formation of the apoptosis-inducing factor (AIF)/phosphorylated H2A histone variant (γ-H2A.X) complex. After mice ingest DEHP, it is metabolized into mono-2-ethylhexyl phthalate (MEHP), which activates Bcl 2-associated X (Bax) to trigger the release of AIF from the mitochondria. The released truncated AIF (tAIF) translocates to the nucleus and forms a DNA degradation complex with γ-H2A.X/cyclophilin A (CypA), leading to DNA fragmentation. In severe cases, this process results in caspase-independent cell death. In contrast, nuclear glutathione peroxidase 4 (nGPx4) overexpression induces chromatin condensation and effectively downregulates γ-H2A.X expression, thereby mitigating caspase-independent cell death. Collectively, these mechanisms protect male mouse germ cells.

Article Snippet: GC-1 cells and mouse testicular tissues were digested and lysed in radioimmunoprecipitation assay (RIPA) buffer (AR0105; Boster) containing protease inhibitors (AR1182; Servicebio) for 30 minutes to extract proteins, including AIF, γ-H2A.X, caspase-3, Bax, and B-cell lymphoma 2 (Bcl-2). nGPx4 and truncated AIF (tAIF) were isolated using a cytoplasmic and nuclear separation kit (AR0106; Boster).

Techniques: Variant Assay, Over Expression, Expressing

Journal: Exploration

Article Title: Tumour‐microenvironment‐responsive Na 2 S 2 O 8 nanocrystals encapsulated in hollow organosilica–metal–phenolic networks for cycling persistent tumour‐dynamic therapy

doi: 10.1002/EXP.20230054

Figure Lengend Snippet: In vitro anticancer activity of Na 2 S 2 O 8 @HTSMSEF. (A) Biocompatibility of HepG2 cells incubated with HTSMSEF for 24 h. (B) Cytotoxicity of HepG2 cells treated by Na 2 S 2 O 8 /Fe 2+ and Na 2 S 2 O 8 @HTSMSEF at various Na 2 S 2 O 8 concentrations ( n = 4). *** p < 0.001 (Student's t ‐test). (C) Flow cytometry plots and (D) cell living/dying of HepG2 cells treated by different formulations. (E) Cell viability of L02 cells (human liver normal cells) and HepG2 cells treated with Na 2 S 2 O 8 /Fe 2+ and Na 2 S 2 O 8 @HTSMSEF at various Na 2 S 2 O 8 concentrations. (F) Confocal images of HepG2 cells treated by different formulations and stained with DCFH‐DA, indicative of intracellular •SO 4 − /•OH radical generation. (G) Confocal images of HepG2 cells treated by different formulations and stained with coumarin‐3‐carboxylic acid, indicative of intracellular •OH generation. (H–J) Confocal images of HepG2 cells treated by different formulations and stained with (H) BCECF‐AM, (I) JC‐1, and (J) γ‐H2AX, indicative of acidification, ΔΨm depolarization, and DNA double‐strand breakage respectively. (K) Confocal images of HepG2 cells treated by different formulations followed by staining with ThiolTracker violet, indicative of intracellular GSH‐depleting ability. (L) Bio‐TEM images of HepG2 cells treated by different formulations.

Article Snippet: After 24 h of culture, the cells were treated with 0.9% NaCl, Na 2 S 2 O 8 /Fe 2+ , HTSMSF, HTSMSEF, Na 2 S 2 O 8 @HTSMSF, and Na 2 S 2 O 8 @HTSMSEF for 8 h. Subsequently, the intracellular DNA double‐strand breakage and ATP level were respectively investigated by using DNA damage assay kit (by γ‐H2AX Immunofluorescence) and Molecular Probes ATP determination assay kit according to the instruction and operation manual.

Techniques: In Vitro, Activity Assay, Incubation, Flow Cytometry, Staining

Journal: Toxicological sciences : an official journal of the Society of Toxicology

Article Title: Comparative Genotoxicity of TEMPO and 3 of Its Derivatives in Mouse Lymphoma Cells

doi: 10.1093/toxsci/kfy022

Figure Lengend Snippet: Criteria for Classification of Chemical’s MoA a

Article Snippet: The genotoxic MoA of TEMPO and its 3 derivatives was investigated by a method for evaluating multiple endpoints that are associated with DNA damage response pathways (MultiFlow DNA Damage Kit—p53, γ-H2AX, Phospho-Histone H3) according to the manufacturer’s manual (Litron Laboratories, Rochester, New York).

Techniques:

Journal: Toxicological sciences : an official journal of the Society of Toxicology

Article Title: Comparative Genotoxicity of TEMPO and 3 of Its Derivatives in Mouse Lymphoma Cells

doi: 10.1093/toxsci/kfy022

Figure Lengend Snippet: MoA of TEMPO and its derivatives evaluated by GEF for multiflow DNA damage assay. Data are expressed as fold-increase over concurrent solvent control and plotted for each chemical. Each dot represents a different concentration of the chemical. Dash line represents the GEF for each endpoint.

Article Snippet: The genotoxic MoA of TEMPO and its 3 derivatives was investigated by a method for evaluating multiple endpoints that are associated with DNA damage response pathways (MultiFlow DNA Damage Kit—p53, γ-H2AX, Phospho-Histone H3) according to the manufacturer’s manual (Litron Laboratories, Rochester, New York).

Techniques: Solvent, Control, Concentration Assay