Journal: Nature Communications

Article Title: Virtual fragment screening for DNA repair inhibitors in vast chemical space

doi: 10.1038/s41467-025-56893-9

Figure Lengend Snippet: a Chemical structures of OGG1 inhibitors TH5487 and compound 23 . b Inhibition of DNA repair enzymes (OGG1, SMUG1, APE1, NEIL1, MTH1, see Methods for abbreviations) by five OGG1 inhibitors. Mean IC 50 values from three independent experiments are shown. Grey boxes indicate IC 50 > 99 μM. c Shift in thermal stabilization of OGG1 in HL60 cells treated with OGG1 inhibitors. Mean values ± SEM from three independent experiments (dot plots) are shown. d Maximum fluorescence intensity of OGG1-GFP accumulation at laser-induced DNA damage sites after indicated treatments. Mean values of 15 cells for each condition from three independent experiments are shown. Statistical significance was determined using One-way Anova ( p = 0.0092 for 10 μM compound 23 , p < 0.0001 for 50 μM compound 23 , and p = 0.0048 for TH5487 ). e Recruitment kinetics of OGG1-GFP to laser-induced DNA damage sites in U2OS cells after 1 h pre-treatment with compound 23 and TH5487 . Results of 15 cells for each condition from three independent experiments are shown ± SEM. f Anti-inflammatory effect of TH5487 and compound 23 in NF-κB activation assay. Mean values ± SD from three independent experiments are shown. g The percentage cell viability of transformed and non-transformed cell lines upon treatment with different OGG1 inhibitors (100 μM). A2780, A549, and HCT116 represent ovarian, lung and colon cancer, respectively, and BJhTERT is a non-transformed control cell line. Mean values from two independent experiments are shown. h Cytotoxicity of TH5487 and compound 11 (Table ) in A2780 and BJhTERT cell lines. Mean values from two independent experiments are shown. Source data are provided as a Source Data file.

Article Snippet: To assess SMUG1 inhibition, a 375 nM substrate containing uracil opposite guanine and 0.3 U SMUG1 enzyme ( M0336 from New England Biolabs) was used.

Techniques: Inhibition, Fluorescence, Activation Assay, Transformation Assay, Control

Journal: Scientific Reports

Article Title: Smug1 alleviates the reproductive toxicity of 5-FU through functioning in rRNA quality control

doi: 10.1038/s41598-025-90330-7

Figure Lengend Snippet: Primer sequences for RT‒qPCR.

Article Snippet: Mouse anti- Smug1 , Santa Cruz , sc-514,343.

Techniques:

Journal: Scientific Reports

Article Title: Smug1 alleviates the reproductive toxicity of 5-FU through functioning in rRNA quality control

doi: 10.1038/s41598-025-90330-7

Figure Lengend Snippet: Antibodies for Western blotting.

Article Snippet: Mouse anti- Smug1 , Santa Cruz , sc-514,343.

Techniques: Western Blot

Journal: Scientific Reports

Article Title: Smug1 alleviates the reproductive toxicity of 5-FU through functioning in rRNA quality control

doi: 10.1038/s41598-025-90330-7

Figure Lengend Snippet: Antibodies for immunofluorescence staining.

Article Snippet: Mouse anti- Smug1 , Santa Cruz , sc-514,343.

Techniques: Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: Smug1 alleviates the reproductive toxicity of 5-FU through functioning in rRNA quality control

doi: 10.1038/s41598-025-90330-7

Figure Lengend Snippet: Expression of Smug1 in oocytes during folliculogenesis and 5-FU-induced changes in Smug1 expression in ovaries and preimplantation embryos. ( a ) Representative immunohistochemistry images showing the expression of Smug1 in the nucleus and cytoplasm of oocytes in different developmental stages of follicles. The arrow indicates the primary-stage oocyte, and Smug1 is presented in brown. Scale bar: 100 μm. ( b ) mRNA expression levels of Smug1 in ovaries obtained from the Ctrl, NR, and R groups were determined via RT‒qPCR. ( c ) Western blot of Smug1 expression in 4-cell-stage embryos in the control group (DMSO) and 5-FU treatment group (500 µM 5-FU). Original blots are presented in Supplementary Fig. 5. ( d ) Quantitation of Smug1 expression in c .

Article Snippet: Mouse anti- Smug1 , Santa Cruz , sc-514,343.

Techniques: Expressing, Immunohistochemistry, Western Blot, Control, Quantitation Assay

Journal: Scientific Reports

Article Title: Smug1 alleviates the reproductive toxicity of 5-FU through functioning in rRNA quality control

doi: 10.1038/s41598-025-90330-7

Figure Lengend Snippet: Knockdown of Smug1 downregulates the nuclear accumulation of Dkc1 and inhibits rRNA maturation, leading to impaired preimplantation embryonic development. ( a ) Representative immunofluorescence images showing the nuclear accumulation of Smug1 and Dkc1 in 4-cell-stage embryos from the Ctrl-MO and Smug1-MO groups. Scale bar: 20 μm. ( b ) Quantitative analysis of the fluorescence intensity of Smug1 (left) and Dkc1 (right). The results were calculated from three independent replicates, and 90 embryos were used for quantification in each group. ( c ) Relative expression levels of 47 S rRNA, 18 S rRNA, 28 S rRNA and 5.8 S rRNA in 4-cell-stage embryos were determined via RT‒qPCR. Zygotes were microinjected with Ctrl-MO (red) or Smug1-MO (purple) and cultured until the 4-cell stage. The data were normalized to the expression of 47 S rRNA in the control group (Ctrl-MO). ( d ) Representative bright-field images of preimplantation embryos at different developmental stages in the Ctrl-MO- and Smug1-MO-injected groups. ( e ) Quantitative graph of preimplantation embryonic development. Ctrl-MO or Smug1-MO was microinjected into zygotes, which were subsequently cultured in vitro. The results were obtained from three independent replicates. A total of 90 or 130 embryos were used for quantification in the Ctrl-MO or Smug1-MO groups, respectively.

Article Snippet: Mouse anti- Smug1 , Santa Cruz , sc-514,343.

Techniques: Knockdown, Immunofluorescence, Fluorescence, Expressing, Cell Culture, Control, Injection, In Vitro

Journal: Scientific Reports

Article Title: Smug1 alleviates the reproductive toxicity of 5-FU through functioning in rRNA quality control

doi: 10.1038/s41598-025-90330-7

Figure Lengend Snippet: Overexpression of Smug1 alleviates the detrimental effects of 5-FU on oocytes and preimplantation embryos by promoting rRNA maturation. ( a ) Representative bright-field images of oocytes microinjected with water or Smug1-flag mRNA, followed by treatment with DMSO or 5-FU. ( b ) Quantitative graph of in vitro maturation rates of control (water injection) or Smug1-overexpressing (Smug1-flag mRNA injection) oocytes that were treated with DMSO or 5-FU. The results were calculated from three independent replicates, and at least 90 oocytes were quantified for each group ( n = 90, effect size: -1.76). ( c ) Representative bright-field images of preimplantation embryos at different developmental stages treated with DMSO or 500 µM 5-FU in the control (water injection) and Smug1-overexpressing (Smug1-flag mRNA injection) groups. ( d ) Quantitative graph of preimplantation embryonic development rates after treatment with DMSO or 500 µM 5-FU in the control and Smug1-overexpressing groups, respectively. The results were calculated from three independent replicates, and at least 90 embryos were quantified for each group. ( e‒g ) Relative expression levels of 47 S rRNA (e), 18 S rRNA (f) and 28 S rRNA (g) in 4-cell-stage embryos determined by RT‒qPCR. DMSO: control group; 500 µM 5-FU: 5-FU treatment group; 500 µM 5-FU + OE-Smug1: Smug1 overexpression combined with 5-FU treatment group.

Article Snippet: Mouse anti- Smug1 , Santa Cruz , sc-514,343.

Techniques: Over Expression, In Vitro, Control, Injection, Expressing

Journal: Scientific Reports

Article Title: Smug1 alleviates the reproductive toxicity of 5-FU through functioning in rRNA quality control

doi: 10.1038/s41598-025-90330-7

Figure Lengend Snippet: LC‒MS/MS analysis of RNA containing 5-Furd. ( a ) RNA Ladder. ( b ) Purified RNA extracted from cells treated with 50 µM 5-FU. ( c ) Purified RNA extracted from Smug1-overexpressing cells treated with 50 µM 5-FU. ( d ) LC‒MS/MS detection chromatogram of standard FUrd, ( e ) FUrd in RNA from cells treated with 50 µM 5-FU, ( f ) 15 N 2 , 13 C-FUrd in RNA from cells treated with 50 µM 5-FU, ( g ) standard FUrd, ( h ) FUrd in RNA extracted from Smug1-overexpressing cells treated with 50 µM 5-FU, ( i ) 15 N 2 , 13 C-FUrd in RNA extracted from Smug1-overexpressing cells treated with 50 µM 5-FU. ( j ) Standard curve. ( k ) Quantitative bar graph of FU content in RNA extracted from control or Smug1-overexpressing cells treated with 50 µM 5-FU. The results were calculated from three independent replicates (effect size: 2.1), 600ng RNA were quantified for each group.

Article Snippet: Mouse anti- Smug1 , Santa Cruz , sc-514,343.

Techniques: Purification, Control