Journal: bioRxiv

Article Title: Sialidases derived from Gardnerella vaginalis remodel the sperm glycocalyx and impair sperm function

doi: 10.1101/2025.02.01.636076

Figure Lengend Snippet: A) Schematic describing lectin flow cytometry and representative glycan epitopes. B, C, D) Sperm were exposed to sialidases (AUS 20 units or GvNanH2 160 units) for 1 hour, fixed in formalin, and stained with MAL-II-biotin/Neutravidin-Alexa Fluor 488, SNA-Cy5, or PSA-FITC respectively. Each point represents the median fluorescence intensity from one donor. Statistics represent a mixed model comparison with post-hoc Holm-Šidák tests. E) Schematic representing change in zeta potential after removal of negatively charged sialic acid on sperm surface. F) Sperm were treated with sialidase at various doses for 1 hour, and zeta potential was measured via dynamic light scattering. Each measurement represents the average of seven technical replicates, and statistics represent a mixed model comparison with post hoc Holm-Šidák tests.

Article Snippet: Biotinylated Maackia Amurensis Lectin II (MAL II, Cat #:B-1265-1) and Cy5-conjugated Sambucus Nigra Lectin (SNA, Cat #: CL-1305-1) were purchased from Vector Labs. Human Contraception Antibody (HCA), was a gift from ZabBio.

Techniques: Flow Cytometry, Staining, Fluorescence, Comparison, Zeta Potential Analyzer

Journal: Experimental and therapeutic medicine

Article Title: Astragaloside IV alleviates heart failure by regulating SUMO-specific protease 1.

doi: 10.3892/etm.2021.10510

Figure Lengend Snippet: Figure 5. Effect of Senp1‑overexpression on H2O2 in HL‑1 cells. (A) The effects of different concentrations of AS‑IV (25, 50, 100 or 200 µmol/l) on the survival of HL‑1 cells subjected to ISO. One‑way ANOVA, followed by Tukey's test. (B) The expression of Senp1 protein was examined by Western blotting following HA‑Senp1 plasmid transfection. T‑test. (C) Compared with the control (dimethyl sulfoxide, 0.1%), H2O2 was increased in the 20 µmol/l ISO‑induced HL‑1 cells. AS‑IV (50 µmol/l, n=6) prevented the ISO‑induced increase in H2O2, which was inhibited by Senp1‑overexpression (n=7). Data are presented as the mean ± standard deviation. Two‑way ANOVA followed by Bonferroni's test. *P<0.05 vs. control; #P<0.05 vs. HF; &P<0.05 vs. HF+AS‑IV. H2O2, hydrogen peroxide; AS‑IV, astragaloside; ANOVA, analysis of variance; ISO, isoprenaline; Senp1, small ubiquitin‑like modifier‑specific protease 1.

Article Snippet: Fetal bovine serum (10099141C) and trypsin (25200072) were purchased from Gibco; Thermo Fisher Scientific, Inc. Antibodies against cleaved‐caspase‐3 (cat. no. 9661, 1:1,000), caspase‐3 (cat. no. 9662, 1:1,000), BCL2 (cat. no. 15071, 1:1,000), Bax (cat. no. 2774, 1:1,000), Senp1 (cat. no. 11929, 1:1,000) and β‐actin (cat. no. 3700, 1:1,000) were purchased from Cell Signaling Technology, Inc.

Techniques: Expressing, Western Blot, Plasmid Preparation, Transfection, Control, Standard Deviation

Journal: Experimental and therapeutic medicine

Article Title: Astragaloside IV alleviates heart failure by regulating SUMO-specific protease 1.

doi: 10.3892/etm.2021.10510

Figure Lengend Snippet: Figure 4. AS‑IV reversed the transverse aortic constriction‑induced expression of apoptosis‑related proteins and Senp1. (A) Apoptosis‑related proteins and Senp1 expression was examined by Western blotting. (B) Quantification of the results in A (n=6). Data are presented as the mean ± standard deviation. One‑way analysis of variance, followed by Tukey's test. *P<0.05 vs. control; #P<0.05 vs. HF. AS‑IV, astragaloside; Senp1, small ubiquitin‑like modifier‑specific protease 1; HF, heart failure; DMSO, dimethyl sulfoxide.

Article Snippet: Fetal bovine serum (10099141C) and trypsin (25200072) were purchased from Gibco; Thermo Fisher Scientific, Inc. Antibodies against cleaved‐caspase‐3 (cat. no. 9661, 1:1,000), caspase‐3 (cat. no. 9662, 1:1,000), BCL2 (cat. no. 15071, 1:1,000), Bax (cat. no. 2774, 1:1,000), Senp1 (cat. no. 11929, 1:1,000) and β‐actin (cat. no. 3700, 1:1,000) were purchased from Cell Signaling Technology, Inc.

Techniques: Expressing, Western Blot, Standard Deviation, Control

Journal: Experimental and therapeutic medicine

Article Title: Astragaloside IV alleviates heart failure by regulating SUMO-specific protease 1.

doi: 10.3892/etm.2021.10510

Figure Lengend Snippet: Figure 7. Effect of Senp1‑overexpression on mitochondrial membrane potential in HL‑1 cells. (A) Compared with the control (dimethyl sulfoxide, 0.1%), the JC‑1 ratio (aggregate/monomer) was decreased in 20 µmol/l ISO‑induced HL‑1 cells. AS‑IV (50 µmol/l, n=7) prevented the ISO‑induced decrease in the JC‑1 ratio, which was inhibited by Senp1‑overexpression (n=8). (B) Summarized data of the JC‑1 ratio. Data are presented as the mean ± standard devia‑ tion. Two‑way analysis of variance followed by Bonferroni's test. *P<0.05 vs. control; #P<0.05 vs. HF; &P<0.05 vs. HF+AS‑IV. Senp1, small ubiquitin‑like modifier‑specific protease 1; ISO, isoprenaline; AS‑IV, astragaloside.

Article Snippet: Fetal bovine serum (10099141C) and trypsin (25200072) were purchased from Gibco; Thermo Fisher Scientific, Inc. Antibodies against cleaved‐caspase‐3 (cat. no. 9661, 1:1,000), caspase‐3 (cat. no. 9662, 1:1,000), BCL2 (cat. no. 15071, 1:1,000), Bax (cat. no. 2774, 1:1,000), Senp1 (cat. no. 11929, 1:1,000) and β‐actin (cat. no. 3700, 1:1,000) were purchased from Cell Signaling Technology, Inc.

Techniques: Membrane, Control

Journal: Experimental and therapeutic medicine

Article Title: Astragaloside IV alleviates heart failure by regulating SUMO-specific protease 1.

doi: 10.3892/etm.2021.10510

Figure Lengend Snippet: Figure 6. Effect of Senp1‑overexpression on ROS generation in HL‑1 cells. (A) Compared with the control (dimethyl sulfoxide, 0.1%), DCF fluorescence was increased in 20 µmol/l ISO‑induced HL‑1 cells. AS‑IV (50 µmol/l, n=6) prevented the ISO‑induced increase in DCF fluorescence, which was inhibited by Senp1‑overexpression. (B) Summarized data of DCF fluorescence (n=7). Data are presented as the mean ± standard deviation. Two‑way analysis of variance, followed by Bonferroni's test. *P<0.05 vs. control; #P<0.05 vs. HF; &P<0.05 vs. HF+AS‑IV. Senp1, small ubiquitin‑like modifier‑specific protease 1; ROS, reac‑ tive oxygen species; ISO, isoprenaline; AS‑IV, astragaloside.

Article Snippet: Fetal bovine serum (10099141C) and trypsin (25200072) were purchased from Gibco; Thermo Fisher Scientific, Inc. Antibodies against cleaved‐caspase‐3 (cat. no. 9661, 1:1,000), caspase‐3 (cat. no. 9662, 1:1,000), BCL2 (cat. no. 15071, 1:1,000), Bax (cat. no. 2774, 1:1,000), Senp1 (cat. no. 11929, 1:1,000) and β‐actin (cat. no. 3700, 1:1,000) were purchased from Cell Signaling Technology, Inc.

Techniques: Control, Fluorescence, Standard Deviation

Journal: Experimental and therapeutic medicine

Article Title: Astragaloside IV alleviates heart failure by regulating SUMO-specific protease 1.

doi: 10.3892/etm.2021.10510

Figure Lengend Snippet: Figure 8. (A) TUNEL staining was used to examine cell apoptosis in HL‑1 cells. Compared with the control (dimethyl sulfoxide, 0.1%), cell apoptosis was increased in 20 µmol/l ISO‑induced HL‑1 cells. AS‑IV (50 µmol/l, n=10) prevented the ISO‑induced increase in cell apoptosis, which was inhibited by Senp1‑overexpression. (B) Summarized data of the cell apoptotic rate (n=7). Two‑way analysis of variance followed by Bonferroni's test. Data are presented as the mean ± standard deviation. *P<0.05 vs. control; #P<0.05 vs. HF; &P<0.05 vs. HF+AS‑IV. ISO, isoprenaline; AS‑IV, astragaloside; Senp1, small ubiquitin‑like modifier‑specific protease 1.

Article Snippet: Fetal bovine serum (10099141C) and trypsin (25200072) were purchased from Gibco; Thermo Fisher Scientific, Inc. Antibodies against cleaved‐caspase‐3 (cat. no. 9661, 1:1,000), caspase‐3 (cat. no. 9662, 1:1,000), BCL2 (cat. no. 15071, 1:1,000), Bax (cat. no. 2774, 1:1,000), Senp1 (cat. no. 11929, 1:1,000) and β‐actin (cat. no. 3700, 1:1,000) were purchased from Cell Signaling Technology, Inc.

Techniques: TUNEL Assay, Staining, Control, Standard Deviation

Journal: International journal of oncology

Article Title: TIP60 governs the auto‑ubiquitination of UHRF1 through USP7 dissociation from the UHRF1/USP7 complex.

doi: 10.3892/ijo.2021.5269

Figure Lengend Snippet: Figure 1. TIP60 and ubiquitin co‑transfection induces the downregulation of UHRF1. Cells were co‑transfected with either TIP60‑eGFP (green) and RFP‑Ubiquitin (red) or eGFP and RFP‑Ubiquitin. Immunostaining of UHRF1 in HeLa cells without (A) or with treatment by MG‑132 (B). Cells were fixed following transfection and labeled with anti‑UHRF1 antibody. Endogenous UHRF1 protein was labeled with Alexa 647‑labeled secondary antibody before visualization with confocal microscopy. Scale bar, 10 µm. (C and D) Represent mean fluorescence intensities levels of UHRF1 in the different samples. Values are the mean ± SEM for three independent experiments; *P<0.05; ****P<0.0001 (vs. control group), determined by one‑way ANOVA with Tukey's post hoc test. UHRF1, ubiquitin‑like, containing PHD and RING finger domains 1; TIP60, Tat interactive protein, 60 kDa.

Article Snippet: Other antibodies used included rabbit polyclonal anti‐HAUSP/USP7 (1:5,000; cat. no. ab4080, Abcam), mouse monoclonal anti‐DNMT1 (1:5,000; cat. no. PTG‐MAB0079, ProteoGenix), mouse monoclonal anti‐ubiquitin (1:500; cat. no. 05‐944, Sigma‐Aldrich; Merck KGaA), mouse monoclonal eGFP (1:1,000; cat. no. 66,002‐1‐Ig, Proteintech Group, Inc.; and cat. no. A‐11120, Thermo Fisher Scientific, Inc.), mouse monoclonal anti‐GAPDH (1:5,000; cat. no. MAB374, Merck KGaA), mouse monoclonal anti‐GFP (1:1,000; cat. no. 66002‐1‐Ig, Proteintech Group, Inc.), mouse mono‐ clonal anti‐p73 (1:500; cat. no. 558785, BD Biosciences), rabbit polyclonal anti‐caspase‐3 (1:1,000; cat. no. 9661, Cell Signaling Technology, Inc.), mouse monoclonal anti‐BCL2 (1:1,000; cat. no. 05‐826, Merck KGaA), mouse monoclonal anti‐poly(ADP‐ribose) polymerase (PARP; 1:1,000; cat. no. 51‐6639GR, BD Biosciences) and rabbit polyclonal anti‐BAX (1:1,000; cat. no. AB2930, Merck KGaA).

Techniques: Ubiquitin Proteomics, Immunostaining, Transfection, Labeling, Confocal Microscopy, Fluorescence, Control

Journal: International journal of oncology

Article Title: TIP60 governs the auto‑ubiquitination of UHRF1 through USP7 dissociation from the UHRF1/USP7 complex.

doi: 10.3892/ijo.2021.5269

Figure Lengend Snippet: Figure 3. TIP60 induces auto‑ubiquitination of UHRF1 in HeLa cells. Cells stably expressing either UHRF1 WT or UHRF1 C724A‑H741A mutant were transfected with either TIP60-eGFP WT or TIP60ΔMYST‑eGFP mutant. All samples were treated with 10 µM of MG‑132, 8 h before harvesting the cells. Whole cell lysates and immunoprecipitated samples were analyzed by SDS‑PAGE and then immunoblotted with anti‑GFP and anti‑Ubiquitin antibodies. Inputs and IP gels were processed in parallel under similar conditions. UHRF1, ubiquitin‑like, containing PHD and RING finger domains 1; TIP60, Tat interactive protein, 60 kDa.

Article Snippet: Other antibodies used included rabbit polyclonal anti‐HAUSP/USP7 (1:5,000; cat. no. ab4080, Abcam), mouse monoclonal anti‐DNMT1 (1:5,000; cat. no. PTG‐MAB0079, ProteoGenix), mouse monoclonal anti‐ubiquitin (1:500; cat. no. 05‐944, Sigma‐Aldrich; Merck KGaA), mouse monoclonal eGFP (1:1,000; cat. no. 66,002‐1‐Ig, Proteintech Group, Inc.; and cat. no. A‐11120, Thermo Fisher Scientific, Inc.), mouse monoclonal anti‐GAPDH (1:5,000; cat. no. MAB374, Merck KGaA), mouse monoclonal anti‐GFP (1:1,000; cat. no. 66002‐1‐Ig, Proteintech Group, Inc.), mouse mono‐ clonal anti‐p73 (1:500; cat. no. 558785, BD Biosciences), rabbit polyclonal anti‐caspase‐3 (1:1,000; cat. no. 9661, Cell Signaling Technology, Inc.), mouse monoclonal anti‐BCL2 (1:1,000; cat. no. 05‐826, Merck KGaA), mouse monoclonal anti‐poly(ADP‐ribose) polymerase (PARP; 1:1,000; cat. no. 51‐6639GR, BD Biosciences) and rabbit polyclonal anti‐BAX (1:1,000; cat. no. AB2930, Merck KGaA).

Techniques: Stable Transfection, Expressing, Mutagenesis, Transfection, Immunoprecipitation