Journal: Cell reports

Article Title: TBK1 is ubiquitinated by TRIM5α to assemble mitophagy machinery

doi: 10.1016/j.celrep.2024.114294

Figure Lengend Snippet: Key resources table

Article Snippet: UBE2N (D2A1) Rabbit mAb , Cell Signaling Technology , #6999, RRID: AB_10828936.

Techniques: Ubiquitin Proteomics, Produced, Purification, Virus, Recombinant, Protease Inhibitor, Lysis, Western Blot, Stripping, Staining, Transfection, Luciferase, Isolation, In Situ, Proximity Ligation Assay, Mutagenesis, Knock-Out, Stable Transfection, Expressing, Plasmid Preparation, Software, Imaging

Journal: Biology Direct

Article Title: UBE2N promotes cell viability and glycolysis by promoting Axin1 ubiquitination in prostate cancer cells

doi: 10.1186/s13062-024-00469-y

Figure Lengend Snippet: Upregulation of UBE2N correlates with the poor prognosis of prostate cancer. (A) UBE2N mRNA is expressed highly in tumor tissues in TCGA-PARD dataset. (B, C) Analysis of UBE2N protein in prostate cancer tissue microarray by IHC staining (scale bar, 100 μm). (D) Survival rate was compared between patients with high and low UBE2N levels by the Log-rank test. (E) UBE2N mRNA levels were examined using RT-qPCR ( n = 3). (F) Western blot determined UBE2N protein expression ( n = 3). Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. normal or HPEPic

Article Snippet: Primary antibodies used in this experiment were as follows: Anti-UBE2N (ab25885; Abcam), anti-Axin1 (#3323; Cell Signaling Technology), anti-c-myc (#5605; Cell Signaling Technology), anti-HK2 (ab104836; Abcam), anti-PKM2 (ab85555; Abcam), anti-Histone H3 (ab1791; Abcam), anti-Ub (ab137031; Abcam), and anti-β-catenin (#8480; Cell Signaling Technology).

Techniques: Microarray, Immunohistochemistry, Quantitative RT-PCR, Western Blot, Expressing

Journal: Biology Direct

Article Title: UBE2N promotes cell viability and glycolysis by promoting Axin1 ubiquitination in prostate cancer cells

doi: 10.1186/s13062-024-00469-y

Figure Lengend Snippet: Knocking down of UBE2N inhibits viability and glycolysis of 22RV1 cells. (A) RT-qPCR and (B) Western blot assay showed UBE2N expression levels in 22RV1 cells transduced with either scramble shRNA (shNC) or shUBE2N-1, -2, and − 3. (C) CCK-8, (D) Extracellular acidification rate (ECAR), (E) oxygen consumption rate (OCR), (F) lactate, (G) ATP production, and (H) expression of c-myc, nuclear β-catenin, HK2 and PKM2. Data are expressed as mean ± SD ( n = 3). *** p < 0.001 vs. shNC

Article Snippet: Primary antibodies used in this experiment were as follows: Anti-UBE2N (ab25885; Abcam), anti-Axin1 (#3323; Cell Signaling Technology), anti-c-myc (#5605; Cell Signaling Technology), anti-HK2 (ab104836; Abcam), anti-PKM2 (ab85555; Abcam), anti-Histone H3 (ab1791; Abcam), anti-Ub (ab137031; Abcam), and anti-β-catenin (#8480; Cell Signaling Technology).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Transduction, shRNA, CCK-8 Assay

Journal: Biology Direct

Article Title: UBE2N promotes cell viability and glycolysis by promoting Axin1 ubiquitination in prostate cancer cells

doi: 10.1186/s13062-024-00469-y

Figure Lengend Snippet: UBE2N knockdown suppresses xenograft growth in a tumor-bearing mouse model. The mouse was injected subcutaneously with 22RV1 cells transduced with either shNC or shUBE2N-1. (A) Tumor volume. On Day 33, xenografts were removed for (B) photographed and (C) weighted. (D, E) PCNA immunofluorescence staining (scale bar, 50 μm). Data are expressed as mean ± SD ( n = 6). * p < 0.05, *** p < 0.001 vs. shNC

Article Snippet: Primary antibodies used in this experiment were as follows: Anti-UBE2N (ab25885; Abcam), anti-Axin1 (#3323; Cell Signaling Technology), anti-c-myc (#5605; Cell Signaling Technology), anti-HK2 (ab104836; Abcam), anti-PKM2 (ab85555; Abcam), anti-Histone H3 (ab1791; Abcam), anti-Ub (ab137031; Abcam), and anti-β-catenin (#8480; Cell Signaling Technology).

Techniques: Injection, Transduction, Immunofluorescence, Staining

Journal: Biology Direct

Article Title: UBE2N promotes cell viability and glycolysis by promoting Axin1 ubiquitination in prostate cancer cells

doi: 10.1186/s13062-024-00469-y

Figure Lengend Snippet: UBE2N overexpression promotes viability and glycolysis in PC3 cells via the Wnt/β-catenin signaling pathway. (A) RT-qPCR and (B) Western blot assay showed UBE2N expression levels in PC3 cells transduced with either UBE2N expression or blank vector. PC3 cells were transduced with either UBE2N expression or blank vector and co-treated with XAV939 or vehicle. (C) Cell viability, (D) ECAR, (E) OCR, (F) lactate, (G) ATP production, and (H) expression of c-myc, nuclear β-catenin, HK2 and PKM2. Data are expressed as mean ± SD ( n = 3). *** p < 0.001 vs. Vector + Vehicle; ### p < 0.001, vs. oeUBE2N + Vehicle

Article Snippet: Primary antibodies used in this experiment were as follows: Anti-UBE2N (ab25885; Abcam), anti-Axin1 (#3323; Cell Signaling Technology), anti-c-myc (#5605; Cell Signaling Technology), anti-HK2 (ab104836; Abcam), anti-PKM2 (ab85555; Abcam), anti-Histone H3 (ab1791; Abcam), anti-Ub (ab137031; Abcam), and anti-β-catenin (#8480; Cell Signaling Technology).

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Expressing, Transduction, Plasmid Preparation

Journal: Biology Direct

Article Title: UBE2N promotes cell viability and glycolysis by promoting Axin1 ubiquitination in prostate cancer cells

doi: 10.1186/s13062-024-00469-y

Figure Lengend Snippet: UBE2N interacts with Axin1 and results in its ubiquitination. (A) Cell lysates were subjected to immunoprecipitation with control IgG, anti-UBE2N, or anti-Axin1 antibody. The immunoprecipitates were then blotted with the indicated antibodies. (B) Axin1 mRNA expression was determined in 22RV1 cells transduced with shUBE2N-1 or -2 and in PC3 cells overexpressing UBE2N. Data are expressed as mean ± SD ( n = 3). (C) Axin1 protein expression was determined in 22RV1 cells transduced with shUBE2N-1 or -2 and in PC3 cells overexpressing UBE2N. (D, E) Axin1 protein expression in PC3 cells overexpressing UBE2N in the absence/presence of MG132 or CHX. (F) 22RV1 cells transduced with shUBE2N-1 or shNC. Axin1 was immunoprecipitated and immunoblotted with the indicated antibodies. (G, H) Analysis of UBE2N protein in prostate cancer tissue microarray by IHC staining (scale bar, 100 μm). (I) Correlation analysis of UBE2N and Axin1 protein expression in prostate cancer tissue microarrays. (J) Axin1 protein levels were examined using Western blot. *** p < 0.001 vs. normal or vector

Article Snippet: Primary antibodies used in this experiment were as follows: Anti-UBE2N (ab25885; Abcam), anti-Axin1 (#3323; Cell Signaling Technology), anti-c-myc (#5605; Cell Signaling Technology), anti-HK2 (ab104836; Abcam), anti-PKM2 (ab85555; Abcam), anti-Histone H3 (ab1791; Abcam), anti-Ub (ab137031; Abcam), and anti-β-catenin (#8480; Cell Signaling Technology).

Techniques: Immunoprecipitation, Expressing, Transduction, Microarray, Immunohistochemistry, Western Blot, Plasmid Preparation

Journal: Biology Direct

Article Title: UBE2N promotes cell viability and glycolysis by promoting Axin1 ubiquitination in prostate cancer cells

doi: 10.1186/s13062-024-00469-y

Figure Lengend Snippet: Axin1 overexpression abrogates the effects of UBE2N overexpression on PC3 cells. (A) RT-qPCR and (B) Western blot assay showed Axin1 expression levels in PC3 cells transfected with Axin1 expression vector or blank vector. PC3 cells were transfected with Axin1 expression vector and transduced with UBE2N expression vector. (C) Cell viability, (D) ECAR, (E) OCR, (F) lactate, (G) ATP production, and (H) expression of c-myc, nuclear β-catenin, HK2 and PKM2. Data are expressed as mean ± SD ( n = 3). *** p < 0.001 vs. Vector + Vector; ### p < 0.001, vs. oeUBE2N + Vector

Article Snippet: Primary antibodies used in this experiment were as follows: Anti-UBE2N (ab25885; Abcam), anti-Axin1 (#3323; Cell Signaling Technology), anti-c-myc (#5605; Cell Signaling Technology), anti-HK2 (ab104836; Abcam), anti-PKM2 (ab85555; Abcam), anti-Histone H3 (ab1791; Abcam), anti-Ub (ab137031; Abcam), and anti-β-catenin (#8480; Cell Signaling Technology).

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Expressing, Transfection, Plasmid Preparation, Transduction

Journal: Biology Direct

Article Title: UBE2N promotes cell viability and glycolysis by promoting Axin1 ubiquitination in prostate cancer cells

doi: 10.1186/s13062-024-00469-y

Figure Lengend Snippet: A diagram of the interaction of UBE2N and Axin1/Wnt/β-catenin signaling in regulating prostate cancer cell viability and glycolysis

Article Snippet: Primary antibodies used in this experiment were as follows: Anti-UBE2N (ab25885; Abcam), anti-Axin1 (#3323; Cell Signaling Technology), anti-c-myc (#5605; Cell Signaling Technology), anti-HK2 (ab104836; Abcam), anti-PKM2 (ab85555; Abcam), anti-Histone H3 (ab1791; Abcam), anti-Ub (ab137031; Abcam), and anti-β-catenin (#8480; Cell Signaling Technology).

Techniques:

Journal: Biology Direct

Article Title: UBE2N promotes cell viability and glycolysis by promoting Axin1 ubiquitination in prostate cancer cells

doi: 10.1186/s13062-024-00469-y

Figure Lengend Snippet: Correlation of tissue high and low levels of UBE2N with clinicopathological features of prostate cancer patients

Article Snippet: Primary antibodies used in this experiment were as follows: Anti-UBE2N (ab25885; Abcam), anti-Axin1 (#3323; Cell Signaling Technology), anti-c-myc (#5605; Cell Signaling Technology), anti-HK2 (ab104836; Abcam), anti-PKM2 (ab85555; Abcam), anti-Histone H3 (ab1791; Abcam), anti-Ub (ab137031; Abcam), and anti-β-catenin (#8480; Cell Signaling Technology).

Techniques: