Journal: Computational and Structural Biotechnology Journal

Article Title: Severe fever with thrombocytopenia syndrome virus (SFTSV)-host interactome screen identifies viral nucleoprotein-associated host factors as potential antiviral targets

doi: 10.1016/j.csbj.2021.09.034

Figure Lengend Snippet: Characterization of the selected SFTSV inhibitors. (A) Dose-response analyses of artenimol and omacetaxine mepesuccinate are shown, depicting both the antiviral activity (red) and cytotoxicity (black). The grey line indicates 50% of the mock-treated control. The EC 50 , CC 50 , and chemical structure of each drug compound are shown. (B) Co-immunoprecipitation assay was conducted in HEK-293T cell transfected with SFTSV nucleoprotein together with either GPI or RPL3 plasmids. After pull-down, the viral nucleoprotein was detected by anti-HA antibody, while GPI and RPL3 were detected by anti-Myc antibodies. (C) Co-immunoprecipitation assay with the treatment of artenimol or omacetaxine mepesuccinate at the indicated concentrations. (D) Molecular docking analysis predicted the interface between artenimol and GPI. The protein is shown as grey ribbons and the drug compound as color sticks. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The plasmids and antibodies, including GPI Human Tagged ORF Clone (RC230292, Origene), RPL3 Human Tagged ORF Clone (RC217987, Origene), mouse anti-HA (Cat #26183; Thermo Fisher), rabbit anti-Myc (Cat #PA1-981, Thermo Fisher), goat anti-mouse-IgG (H + L) HRP (A16072SAMPLE, Thermo Fisher), and goat anti-rabbit-IgG (H + L) HRP (A16104SAMPLE, Thermo Fisher) were purchased from commercial sources.

Techniques: Activity Assay, Control, Co-Immunoprecipitation Assay, Transfection

Journal: Journal of clinical pathology

Article Title: Immunohistochemical panel to differentiate endometrial stromal sarcoma, uterine leiomyosarcoma and leiomyoma: something old and something new

doi: 10.1136/jclinpath-2015-202915

Figure Lengend Snippet: Conditions and titrations of the 10 antibodies

Article Snippet: ASC1 , Novus , 1:100 , PH 8.0 20 min , liver.

Techniques: Control

Journal: Journal of clinical pathology

Article Title: Immunohistochemical panel to differentiate endometrial stromal sarcoma, uterine leiomyosarcoma and leiomyoma: something old and something new

doi: 10.1136/jclinpath-2015-202915

Figure Lengend Snippet: Expressions of the 10 antibodies in 94 cases of leiomyoma, ESS and ULMS

Article Snippet: ASC1 , Novus , 1:100 , PH 8.0 20 min , liver.

Techniques:

Journal: Journal of clinical pathology

Article Title: Immunohistochemical panel to differentiate endometrial stromal sarcoma, uterine leiomyosarcoma and leiomyoma: something old and something new

doi: 10.1136/jclinpath-2015-202915

Figure Lengend Snippet: Positive expression of ASC1 in (A) low grade (LG) endometrial stromal sarcoma (ESS), (B) ‘LG’ uterine leiomyosarcoma (ULMS), (C) leiomyoma (LM). ASC1 was strongly expressed in cytoplasmic and nuclear patterns. Positive expression of stathmin1 in (D) in ESS, (E) leiomyosarcoma and (F) LM. Stathmin1 was strongly expressed in cytoplasmic and nuclear patterns.

Article Snippet: ASC1 , Novus , 1:100 , PH 8.0 20 min , liver.

Techniques: Expressing

Journal: Journal of clinical pathology

Article Title: Immunohistochemical panel to differentiate endometrial stromal sarcoma, uterine leiomyosarcoma and leiomyoma: something old and something new

doi: 10.1136/jclinpath-2015-202915

Figure Lengend Snippet: Comparison of biomarker expressions between all ESS and ULMS

Article Snippet: ASC1 , Novus , 1:100 , PH 8.0 20 min , liver.

Techniques: Comparison, Biomarker Discovery

Journal: Journal of clinical pathology

Article Title: Immunohistochemical panel to differentiate endometrial stromal sarcoma, uterine leiomyosarcoma and leiomyoma: something old and something new

doi: 10.1136/jclinpath-2015-202915

Figure Lengend Snippet: Comparison of biomarker expressions between LG ESS and ‘LG’ ULMS

Article Snippet: ASC1 , Novus , 1:100 , PH 8.0 20 min , liver.

Techniques: Comparison, Biomarker Discovery

Journal: Molecular Cell

Article Title: The ASC-1 Complex Disassembles Collided Ribosomes

doi: 10.1016/j.molcel.2020.06.006

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-ASC-1 , Bethyl , Cat. #A300-843A.

Techniques: Recombinant, Protease Inhibitor, Plasmid Preparation, Sequencing, Mutagenesis, Software, Cloning

Journal: Biomarker Research

Article Title: Histone modification-regulated LncRNA DLEU1 interacts with ASCC2/ALKBH3 complex to drive DNA repair, antioxidant homeostasis and glucose metabolism in gastric cancer

doi: 10.1186/s40364-025-00867-y

Figure Lengend Snippet: DLEU1 facilitates ASCC2 nuclear translocation and promotes ASCC2–ALKBH3 interaction. A . Silver staining (left panel) analysis of proteins isolated from AGS cells using ChIRP with DLEU1 or the control LacZ probes. The red rectangle highlighted the potential presence of ASCC2 in the DLEU1 complex, subsequently confirmed by LC/MS mass spectrometry analysis (right panel). B - C . RNA pull-down ( B ) and RIP assays ( C ) demonstrated the interaction between ASCC2 and DLEU1 in AGS and HGC27 cells. D . Immunofluorescence staining showed co-localization of DLEU1 with ASCC2, Scale bar = 10 μm. E . Western blot results showed that overexpression of DLEU1 increased ASCC2 protein levels in MGC803 and AGS cells, whereas knockdown of DLEU1 decreased ASCC2 protein levels in HGC27 and MKN45 cells. F - G . Nuclear-cytoplasmic fractions ( F ) and Immunofluorescence staining ( G ) demonstrate increased nuclear localization of ASCC2 in DLEU1 overexpressed cells. Scale bar = 10 μm. H . Endogenous co-immunoprecipitation assays reveal enhanced ASCC2–ALKBH3 binding following DLEU1 overexpression. I . Western blot analysis showed changes in ASCC2 and ALKBH3 protein expression in AGS with DLEU1 overexpression or combined DLEU1 overexpression with ASCC2 knockdown(left panel), and in HGC27 cells with DLEU1 knockdown or combined DLEU1 knockdown with ASCC2 overexpression (right panel). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant. See also Figures

Article Snippet: The staining sequence and dye combination are as follows: DLEU1 probe ( NR_109973.1 , 5-1627, working solution, Alpha X Bio, China) - TSA520; E2F1 (1/200 dilution, Proteintech, Cat. #66515-1-Ig, RRID: AB_2881878) - TSA480; ALKBH3 (1/200 dilution, Proteintech, Cat. #12292-1-AP, RRID: AB_11125161) - TSA620; and ASCC2 (1/200 dilution, Proteintech, Cat.#11529-1-AP, RRID: AB_2059358) - TSA780.

Techniques: Translocation Assay, Silver Staining, Isolation, Control, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Immunofluorescence, Staining, Western Blot, Over Expression, Knockdown, Immunoprecipitation, Binding Assay, Expressing

Journal: Biomarker Research

Article Title: Histone modification-regulated LncRNA DLEU1 interacts with ASCC2/ALKBH3 complex to drive DNA repair, antioxidant homeostasis and glucose metabolism in gastric cancer

doi: 10.1186/s40364-025-00867-y

Figure Lengend Snippet: DLEU1 promotes G6PD transcription via ASCC2/ALKBH3-mediated recruitment of E2F1. A - B . qRT-PCR and Western blot analysis of E2F1 expression in AGS and MGC803 cells upon ALKBH3 overexpression. C . Time-course analysis of E2F1 mRNA levels in AGS and MGC803 cells transfected with vector or ALKBH3. D - E . RT-qPCR and western blot analyzed of E2F1 and G6PD expression in AGS and MGC803 cells following DLEU1 overexpression or DLEU1 overexpression combined with ASCC2 knockdown. F - G . RT-qPCR and western blot analyzed of G6PD levels in AGS and MGC803 cells with E2F1 overexpression. H . qRT-PCR analysis of G6PD mRNA expression in AGS and MGC803 cells after ALKBH3 overexpression or co-transfection with ALKBH3 and shRNA targeting E2F1 (shE2F1). I . Western blot analysis of ASCC2, G6PD and E2F1 expression in AGS and MGC803 cells with ALKBH3 overexpression or ALKBH3 overexpression combined with E2F1 knockdown. J . ChIP-qPCR analysis confirmed E2F1 binding to the G6PD promoter and intron 2 regions in AGS cells. K . Luciferase reporter assay showed the transcriptional activity of wild-type and mutant G6PD promoter and intron 2 regions in AGS cells. L . Luciferase assays demonstrate that knockdown of DLEU1 , ASCC2, or ALKBH3 attenuates E2F1-mediated activation of G6PD transcription. L - M . ChIP-qPCR and luciferase reporter validated E2F1 binding to an intronic region of ASCC2 in AGS cells. M . Luciferase assays indicate that knockdown of DLEU1 , ASCC2, or ALKBH3 reduces E2F1-driven ASCC2 transcription. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant

Article Snippet: The staining sequence and dye combination are as follows: DLEU1 probe ( NR_109973.1 , 5-1627, working solution, Alpha X Bio, China) - TSA520; E2F1 (1/200 dilution, Proteintech, Cat. #66515-1-Ig, RRID: AB_2881878) - TSA480; ALKBH3 (1/200 dilution, Proteintech, Cat. #12292-1-AP, RRID: AB_11125161) - TSA620; and ASCC2 (1/200 dilution, Proteintech, Cat.#11529-1-AP, RRID: AB_2059358) - TSA780.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Over Expression, Transfection, Plasmid Preparation, Knockdown, Cotransfection, shRNA, ChIP-qPCR, Binding Assay, Luciferase, Reporter Assay, Activity Assay, Mutagenesis, Activation Assay

Journal: Biomarker Research

Article Title: Histone modification-regulated LncRNA DLEU1 interacts with ASCC2/ALKBH3 complex to drive DNA repair, antioxidant homeostasis and glucose metabolism in gastric cancer

doi: 10.1186/s40364-025-00867-y

Figure Lengend Snippet: Inhibition of ASCC2 abrogates DLEU1 -mediated DNA damage repair and apoptosis resistance in GC cells. A . Proliferation assays in AGS and MGC803 with DLEU1 overexpression, with or without ASCC2 knockdown, and in HGC27 and MKN45 cells with DLEU1 knockdown, with or without ASCC2 overexpression. B - C . Comet assay, Western blot analysis of γ-H2AX protein with DLEU1 overexpression or DLEU1 overexpression combined with ASCC2 knockdown. Scale bar = 50 μm. D - E . Immunofluorescence staining of γH2AX in AGS cells treated with Etoposide. Representative images at 0, 4, 8, and 12 h after treatment are shown. Quantification of average foci number is shown in figure E . Scale bar = 100 μm. F . Apoptosis analysis by flow cytometry in AGS and MGC803 with DLEU1 overexpression or DLEU1 overexpression combined with ASCC2 knockdown. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant

Article Snippet: The staining sequence and dye combination are as follows: DLEU1 probe ( NR_109973.1 , 5-1627, working solution, Alpha X Bio, China) - TSA520; E2F1 (1/200 dilution, Proteintech, Cat. #66515-1-Ig, RRID: AB_2881878) - TSA480; ALKBH3 (1/200 dilution, Proteintech, Cat. #12292-1-AP, RRID: AB_11125161) - TSA620; and ASCC2 (1/200 dilution, Proteintech, Cat.#11529-1-AP, RRID: AB_2059358) - TSA780.

Techniques: Inhibition, Over Expression, Knockdown, Single Cell Gel Electrophoresis, Western Blot, Immunofluorescence, Staining, Flow Cytometry

Journal: Biomarker Research

Article Title: Histone modification-regulated LncRNA DLEU1 interacts with ASCC2/ALKBH3 complex to drive DNA repair, antioxidant homeostasis and glucose metabolism in gastric cancer

doi: 10.1186/s40364-025-00867-y

Figure Lengend Snippet: DLEU1 knockdown suppresses tumor growth by targeting ASCC2 and G6PD in xenograft models. A . Schematic representation of the xenograft experiment. MKN45 cells with stable knockdown of DLEU1 ( shDLEU1 ) or co-knockdown of DLEU1 and ASCC2 ( shDLEU1 + shASCC2 ) were subcutaneously implanted into nude mice. G6PD inhibitor (G6PDi-1) was administered intraperitoneally (10 mg/kg, every 2 days, for a total of six doses) from day 15 to day 21. B . Tumor growth analysis across different groups. Left: Representative images of tumors from the indicated groups. Middle: Tumor weight comparison among groups. Right: Tumor volume progression over time. C . IHC staining for ASCC2, ALKBH3, E2F1, and G6PD in tumor tissues. Representative IHC images (left) and quantification of staining scores (right) for each marker. Scale bar = 50 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant

Article Snippet: The staining sequence and dye combination are as follows: DLEU1 probe ( NR_109973.1 , 5-1627, working solution, Alpha X Bio, China) - TSA520; E2F1 (1/200 dilution, Proteintech, Cat. #66515-1-Ig, RRID: AB_2881878) - TSA480; ALKBH3 (1/200 dilution, Proteintech, Cat. #12292-1-AP, RRID: AB_11125161) - TSA620; and ASCC2 (1/200 dilution, Proteintech, Cat.#11529-1-AP, RRID: AB_2059358) - TSA780.

Techniques: Knockdown, Comparison, Immunohistochemistry, Staining, Marker

Journal: Biomarker Research

Article Title: Histone modification-regulated LncRNA DLEU1 interacts with ASCC2/ALKBH3 complex to drive DNA repair, antioxidant homeostasis and glucose metabolism in gastric cancer

doi: 10.1186/s40364-025-00867-y

Figure Lengend Snippet: Expression and correlation analysis of DLEU1 and associated markers in normal and GC tissues, and its epigenetic regulation in GC. A . Representative multiplex immunofluorescence staining images showing DLEU1 (green), ASCC2 (white), ALKBH3 (orange), and E2F1 (cyan) in normal and tumor tissues ( N = 26, T = 104). Scale bar = 50 μm. B . Quantification of DLEU1 -positive cell density in normal and tumor tissues. C . Quantification of ASCC2-, ALKBH3-, and E2F1-positive cell densities in normal and tumor tissues. D . Percentage of E2F1 localization in the nucleus and cytoplasm in normal and tumor tissues. E . Scatter plots showing positive correlations between DLEU1 expression and ASCC2, ALKBH3, and E2F1 expression in tumor tissues. F . Correlation between DLEU1 expression and E2F1 localization in the cytoplasm and nucleus in tumor tissues. G . Correlation between ASCC2- and ALKBH3-positive cell in tumor tissues. H . HGC27 and MKN45 cells were treated with DMSO (control), Etoposide (10 µM, 24 h), 5-Fluorouracil (5-FU, 20 µg/mL, 24 h), or H 2 O 2 (10 µM, 2 h). The enrichment of H3K4me3 and H3K27ac at the DLEU1 promoter was analyzed by ChIP-qPCR. I . Proposed model illustrating how DLEU1 promotes gastric cancer progression via the ASCC2/ALKBH3/E2F1/G6PD axis, impacting transcriptional activation, DNA repair, and redox homeostasis. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant

Article Snippet: The staining sequence and dye combination are as follows: DLEU1 probe ( NR_109973.1 , 5-1627, working solution, Alpha X Bio, China) - TSA520; E2F1 (1/200 dilution, Proteintech, Cat. #66515-1-Ig, RRID: AB_2881878) - TSA480; ALKBH3 (1/200 dilution, Proteintech, Cat. #12292-1-AP, RRID: AB_11125161) - TSA620; and ASCC2 (1/200 dilution, Proteintech, Cat.#11529-1-AP, RRID: AB_2059358) - TSA780.

Techniques: Expressing, Multiplex Assay, Immunofluorescence, Staining, Control, ChIP-qPCR, Activation Assay

Journal: Cancers

Article Title: N-Acetylcysteine Promotes Metastatic Spread of Melanoma in Mice

doi: 10.3390/cancers14153614

Figure Lengend Snippet: Effect of inhibition of Cys/cystine uptake or GSH synthesis on GSH levels and metastatic activity in B16-F10 cells. Cancer cells were transfected in vitro, as explained under Material and Methods, before their in vivo inoculation. B16-F10 cells were inoculated i.v. (portal vein). NAC (240 mg/kg) was administered orally for 10 days, starting 120 min after B16-F10 cells inoculation. All measurements were performed in the liver or in metastatic cells isolated from the liver 10 days after tumor inoculation. Results obtained using melanoma cells treated with scrambled RNA sequences were not significantly different from those displayed as non-treated controls. * Significantly different p < 0.01, comparing shRNA-treated cells versus controls; + Significantly different p < 0.01, comparing NAC-treated mice versus mice treated with vehicle (n = 9–10).

Article Snippet: Knockdown of SLC7A10 (codes the ASC1 Na + -independent neutral amino acid transporter) was performed using an ASC1 shRNA Plasmid sc-39159-SH from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Inhibition, Activity Assay, Transfection, In Vitro, In Vivo, Isolation, shRNA, Control