Journal: Nature Communications

Article Title: Targeting USP47 overcomes tyrosine kinase inhibitor resistance and eradicates leukemia stem/progenitor cells in chronic myelogenous leukemia

doi: 10.1038/s41467-020-20259-0

Figure Lengend Snippet: a YB-1 was silenced in K562, K562R, and KBM5 T315I cells with a retroviral transduction system, and viable cells were counted in the transfected cell lines at different times ( n = 3 biologically independent samples per group). Data are mean ± s.d. p -values were analyzed by two-way analysis of variance (ANOVA). **** p < 0.0001. b DNA damage-related protein expression was measured by western blot after USP47 knockdown in the K562 and KBM5 T315I cells. c GFP + BM mononuclear cells were collected from Usp47 +/+ and Usp47 −/− CML mice, and the indicated proteins were examined by western blot. d Immunofluorescence staining of γH 2 AX foci in the USP47 or YB-1 knockdown K562 cells. Scale bars, 20 μm. e The number of AP sites (DNA damage quantification) was detected in genomic DNA after USP47 knockdown in the K562 and KBM5 T315I cells ( n = 3 biologically independent samples per group). Data are mean ± s.d. p -values were analyzed by one-way analysis of variance (ANOVA). *** p < 0.001. f Time-course analysis of the USP47 knockdown-induced DNA damage response in K562 cells and cleaved-PARP1 by western blot. g YB-1 , PCNA , and TOPO IIα mRNA levels after YB-1 knockdown at day 7 in K562 cells ( n = 3 biologically independent samples per group). Data are mean ± s.d. p- values were analyzed by one-way analysis of variance (ANOVA). * p < 0.05; ** p < 0.01; **** p < 0.0001. h DNA damage-related protein expression was measured by western blot after YB-1 knockdown in the K562 cells. i The time-course of DNA damage is shown by γH 2 AX expression after irradiation (IR, 3 Gy) in the control and YB-1 stably knockdown KBM5 T315I cells. Band intensity (γH 2 AX relative to β-actin) is shown by the histogram ( n = 3 biologically independent samples per group). Data are mean ± s.d. p- values were analyzed by two-way analysis of variance (ANOVA). *** p < 0.001; **** p < 0.0001. j Exogenous YB-1 protein was ex p ressed in the cell line with stably knocked down USP47 , the expression of γH 2 AX and exogenous YB-1 (GFP tag) was detected by western blot. k The number of AP sites was measured as described above ( n = 3 biologically independent samples per group). Data are mean ± s.d. p- values were analyzed by one-way analysis of variance (ANOVA). *** p < 0.001. l γH 2 AX expression in Usp47 −/− MEFs was measured by western blot after the reintroduction of YB-1. m , n USP47 was knocked down in primary CML CD34 + cells, then the cells were cultured in a stem cell colony formation medium. The colonies ( m ) were counted on day 14 ( n = 3 biologically independent samples per group). Data are mean ± s.d. p- values were analyzed by one-way analysis of variance (ANOVA). *** p < 0.001. Immunofluorescence staining of γH 2 AX foci ( n ) was performed ( n = 3 biologically independent samples per group). Scale bars, 20 μm. Source data are provided as a Source Data file.

Article Snippet: The number of apurinic/apyrimidinic sites (AP sites) was determined using the DNA Damage Quantification Kit (DK02, DOJINDO Laboratories, Kumamoto, Japan) according to the manufacturer’s instruction.

Techniques: Retroviral, Transduction, Transfection, Expressing, Western Blot, Knockdown, Immunofluorescence, Staining, Irradiation, Control, Stable Transfection, Cell Culture

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: Antioxidants (Basel, Switzerland)

Article Title: Oral Curcumin-Thioketal-Inulin Conjugate Micelles against Radiation-Induced Enteritis.

doi: 10.3390/antiox13040417

Figure Lengend Snippet: Figure 5. CTI protects intestinal cells from radiation-induced damage. (A) Cell viability of IEC- 6 cells after different treatments. (B) Flow cytometry plots of IEC-6 cells after FITC–PI staining. (C) Population of cell apoptosis after radiation. (D) Calcein AM–PI dyeing of IEC-6 cells after different treatments. (E) Representative photographs of colony numbers with different treatments. (F) Quantitative analysis of the percentage of colony numbers after different treatments. (G) ROS levels were measured with a fluorescence microplate reader. (H) Intracellular ROS measurement of L929 cells by DCFH-DA staining (scale bars, 100 µm). (I) DNA double-strand damage in IEC-6 cells after different treatments by γH2AX immunofluorescence assay. Data are presented as means ± SD, n = 3 (** p < 0.01, *** p < 0.001, n = 3).

Article Snippet: After attachment, cells were treated with 10 μg/mL of Cur or 100 μg/mL of CTI micelles for 4 h. Then, 24 h after irradiation, the cells were fixed with 4% paraformaldehyde for 20 min and washed with PBS and treated with 0.2% Triton-X 100 for 15 min. After PBS washing, the cells were incubated with rabbit polyclonal γH2AX (phospho S139) primary antibody (dilution 1:1000; G266; Dojindo, Tokyo, Japan) for 1 h. After discarding the primary antibody, the cells were gently washed with PBS three times and incubated with goat anti-rabbit secondary antibody (G266; Dojindo) at room temperature for 1 h. The nuclei were counterstained with DAPI (C0065, Solarbio, Beijing, China).

Techniques: Flow Cytometry, Staining, Fluorescence, Immunofluorescence

Journal: Experimental and Therapeutic Medicine

Article Title: Circulating levels of DDIT4 and mTOR, and contributions of BMI, inflammation and insulin sensitivity in hyperlipidemia

doi: 10.3892/etm.2022.11602

Figure Lengend Snippet: Comparisons of DDIT4, mTOR, and inflammatory factors between the two groups.

Article Snippet: The serum concentrations of DDIT4 (cat. no. CSB-EL006590HU) and mTOR (cat. no. CSB-E09038h) were measured using ELISA kits from Cusabio according to manufacturer's protocol.

Techniques:

Journal: Experimental and Therapeutic Medicine

Article Title: Circulating levels of DDIT4 and mTOR, and contributions of BMI, inflammation and insulin sensitivity in hyperlipidemia

doi: 10.3892/etm.2022.11602

Figure Lengend Snippet: Correlations among clinical data and metabolic and inflammatory parameters.

Article Snippet: The serum concentrations of DDIT4 (cat. no. CSB-EL006590HU) and mTOR (cat. no. CSB-E09038h) were measured using ELISA kits from Cusabio according to manufacturer's protocol.

Techniques:

Journal: Experimental and Therapeutic Medicine

Article Title: Circulating levels of DDIT4 and mTOR, and contributions of BMI, inflammation and insulin sensitivity in hyperlipidemia

doi: 10.3892/etm.2022.11602

Figure Lengend Snippet: Results of the principal component factor analysis with a Varimax rotation among subjects with hyperlipidemia.

Article Snippet: The serum concentrations of DDIT4 (cat. no. CSB-EL006590HU) and mTOR (cat. no. CSB-E09038h) were measured using ELISA kits from Cusabio according to manufacturer's protocol.

Techniques:

Journal: Animals : an Open Access Journal from MDPI

Article Title: Assessing Allostatic Load in Ring-Tailed Lemurs ( Lemur catta )

doi: 10.3390/ani11113074

Figure Lengend Snippet: Biomarkers used in the estimation of allostatic load for in ring-tailed lemurs ( Lemur catta ) and the response of the biomarkers to chronic stress.

Article Snippet: The DNA damage ELISA is a competitive monoclonal assay designed to measure RNA and DNA oxidized guanine species (DetectX ® DNA Damage Immunoassay Catalog # K059-H1, Arbor Assays Interactive Assay Solutions, Ann Arbor, MI, USA).

Techniques: Functional Assay