Journal: Cell Death & Disease

Article Title: FBXO6-mediated RNASET2 ubiquitination and degradation governs the development of ovarian cancer

doi: 10.1038/s41419-021-03580-4

Figure Lengend Snippet: A 293T cells were co-transfected with Flag-FBXO6 and HA-RNASET2 for 36 h. Cells were collected and lysed. The WCLs were immunoprecipitated by anti-Flag M2 resin and subjected to western blot analysis as indicated. B A2780 cells were treated with MG132 for 4 h. Cells were then collected and lysed. Lysates were immunoprecipitated with anti-FBXO6 antibody and analyzed by immunoblotting as indicated. A nonspecific IgG was used as a negative control. C OVCAR-3 cells were treated with MG132 for 4 h. Cells were then collected and lysed. Lysates were immunoprecipitated with anti-RNASET2 antibody and analyzed by immunoblotting as indicated. A nonspecific IgG was used as a negative control. D 293T cells were transfected with Flag-Con or the indicated FLAG-tagged F-box protein plasmids (FBPs) for 36 h. Cells were collected and lysed. The WCLs were immunoprecipitated by anti-Flag M2 resin, eluted by 3× Flag peptides and immunoblotting as indicated. E The figure represents the domains of the FBXO6 deletion constructs. F 293T cells were transfected with the indicated Flag-tagged FBXO6 deletion constructs for 36 h. Cells were collected and lysed. The WCLs were immunoprecipitated by anti-Flag M2 resin, eluted by 3× Flag peptides and immunoblotting as indicated. G OVCAR-3 cells were transfected with the Flag-FBXO6 WT or Flag-FBXO6 Mut for 36 h. Cells were collected and lysed. The WCLs were immunoprecipitated by anti-Flag M2 resin, eluted by 3× Flag peptides and immunoblotting as indicated.

Article Snippet: A2780 cells were transfected with FBXO6 CRISPR/Cas9 KO (h) KO plasmid (sc-405919, Santa Cruz Biotechnology) using Lipofectamine 2000 following the manufacturer’s instructions.

Techniques: Transfection, Immunoprecipitation, Western Blot, Negative Control, Construct

Journal: Cell Death & Disease

Article Title: FBXO6-mediated RNASET2 ubiquitination and degradation governs the development of ovarian cancer

doi: 10.1038/s41419-021-03580-4

Figure Lengend Snippet: A Western blot analysis of OVCAR-3 cells transfected with Flag-Con or increased Flag-FBXO6 plasmids. B Western blot analysis of OVCAR-3 cells transfected with scramble or FBXO6 shRNAs. C A2780 FBXO6 knockout (FBXO6 −/− ) cells were generated by CRISPR assay and the WCLs were subjected to western blot analysis with indicated antibodies. D The mRNA levels of RNASET2 in C were determined by real-time PCR assay. E FBXO6 +/+ and FBXO6 −/− A2780 cells were treated with 50 μg/mL CHX for the indicated time. Cells were lysed and the WCLs were subjected to western blot analysis with the indicated antibodies. Statistic results of immunoblotting analysis were obtained by ImageJ software and were normalized to GAPDH intensities. Error bars indicate the means ± SD, n = 3. F FBXO6 +/+ and FBXO6 −/− A2780 cells were treated with or without 10 μM MG132 for 4 h. Cells were lysed and the WCLs were subjected to western blot analysis with the indicated antibodies. G A2780 cells were transfected with con-shRNA or FBXO6-shRNA for 72 h. Cells were then either untreated or transfected with Flag-Con, FBXO6 WT, and FBXO6 Mut for an additional 48 h, respectively. Cells were lysed and the WCLs were subjected to western blot analysis with the indicated antibodies. H 293T cells were transfected with HA-RNASET2 with or without Flag-FBXO6 and His-ubiquitin for 36 h. Cells were lysed and the WCLs were immunoprecipitated by anti-HA resin and immunoblotting as indicated.

Article Snippet: A2780 cells were transfected with FBXO6 CRISPR/Cas9 KO (h) KO plasmid (sc-405919, Santa Cruz Biotechnology) using Lipofectamine 2000 following the manufacturer’s instructions.

Techniques: Western Blot, Transfection, Knock-Out, Generated, CRISPR, Real-time Polymerase Chain Reaction, Software, shRNA, Ubiquitin Proteomics, Immunoprecipitation

Journal: Cell Death & Disease

Article Title: FBXO6-mediated RNASET2 ubiquitination and degradation governs the development of ovarian cancer

doi: 10.1038/s41419-021-03580-4

Figure Lengend Snippet: A The cell growth curve of OVCAR-3 cells transfected with scramble or FBXO6 shRNAs. B Clonogenic assay of OVCAR-3 cells transfected with scramble or FBXO6 shRNAs. C The cell growth curve of FBXO6 +/+ and FBXO6 −/− A2780 cells. D Clonogenic assay of FBXO6 +/+ and FBXO6 −/− A2780 cells. E Representative images of migrated FBXO6 +/+ and FBXO6 − /− A2780 cells in a transwell assay with Matrigel. F Quantification of migrated cells in E . Data were shown as mean ± SD of three independent experiments. ** p < 0.01. G The migration of FBXO6 +/+ and FBXO6 −/− A2780 cells in scratch experiments. Data were shown as mean ± SD of three independent experiments. ** p < 0.01. H FBXO6 +/+ (1 × 10 7 ) and FBXO6 −/− A2780 cells were subcutaneously injected into each nude mouse for about 4 weeks. Tumor volumes were detected at the indicated dates. n = 6 for each group. *** p < 0.001. I The image shows the representative tumor-bearing mice and xenografts for each group. J At the end of 4 weeks, mice were killed and the tumor weights were measured. ** p < 0.01.

Article Snippet: A2780 cells were transfected with FBXO6 CRISPR/Cas9 KO (h) KO plasmid (sc-405919, Santa Cruz Biotechnology) using Lipofectamine 2000 following the manufacturer’s instructions.

Techniques: Transfection, Clonogenic Assay, Transwell Assay, Migration, Injection

Journal: Cell Death & Disease

Article Title: FBXO6-mediated RNASET2 ubiquitination and degradation governs the development of ovarian cancer

doi: 10.1038/s41419-021-03580-4

Figure Lengend Snippet: A The mRNA expression of FBXO6 between 426 ovarian cancer tissues and 88 non‐tumor tissues in TCGA database. B The overall survival (OS) curve of FBXO6 gene expression in ovarian cancer patients with advanced stages ( http://kmplot.com/analysis/ ). C Immunohistochemical analyses of 88 specimens from ovarian cancer patients using anti-FBXO6 and anti-RNASET2 antibodies were performed. Representative images of IHC staining of tumors from two human ovarian cancer patients are presented. D The correlation study of FBXO6 and RNASET2 protein expression in 88 ovarian cancer samples is shown.

Article Snippet: A2780 cells were transfected with FBXO6 CRISPR/Cas9 KO (h) KO plasmid (sc-405919, Santa Cruz Biotechnology) using Lipofectamine 2000 following the manufacturer’s instructions.

Techniques: Expressing, Gene Expression, Immunohistochemical staining, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: CD8+ T Cell Co-Expressed Genes Correlate With Clinical Phenotype and Microenvironments of Urothelial Cancer

doi: 10.3389/fonc.2020.553399

Figure Lengend Snippet: The Module and Gene significance for CD8 + T Cells related genes.

Article Snippet: The proteins level encoded by these genes ( PSMB10 , PSMB9 , PSMB8 , TAP1 , IRF1 , and FBXO6 ) were lower in the high clinical grade patients in the human protein atlas (HPA), which suggested the clinical phenotype correlation both in mRNA and protein levels ( ).

Techniques:

Journal: Frontiers in Oncology

Article Title: CD8+ T Cell Co-Expressed Genes Correlate With Clinical Phenotype and Microenvironments of Urothelial Cancer

doi: 10.3389/fonc.2020.553399

Figure Lengend Snippet: Univariate Cox proportional hazard analysis of CD8 + T cells related genes.

Article Snippet: The proteins level encoded by these genes ( PSMB10 , PSMB9 , PSMB8 , TAP1 , IRF1 , and FBXO6 ) were lower in the high clinical grade patients in the human protein atlas (HPA), which suggested the clinical phenotype correlation both in mRNA and protein levels ( ).

Techniques:

Journal: Frontiers in Oncology

Article Title: CD8+ T Cell Co-Expressed Genes Correlate With Clinical Phenotype and Microenvironments of Urothelial Cancer

doi: 10.3389/fonc.2020.553399

Figure Lengend Snippet: (A) The proteins level encoded by these genes ( PSMB10 , PSMB9 , PSMB8 , TAP1 , IRF1 , and FBXO6 ) were lower in the high clinical grade patients in the human protein atlas (HPA). (B) The results of GSEA analysis. Antigen processing and presentation, the chemokine signaling pathway, nature killer cell-mediated cytotoxicity, and the T cell receptor signaling pathway were related to the high expression group in ETV7 , FBXO6 , IRF1 , PSMB8 , PSMB9 , PSMB10 , PSME2 , and TAP1 .

Article Snippet: The proteins level encoded by these genes ( PSMB10 , PSMB9 , PSMB8 , TAP1 , IRF1 , and FBXO6 ) were lower in the high clinical grade patients in the human protein atlas (HPA), which suggested the clinical phenotype correlation both in mRNA and protein levels ( ).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: CD8+ T Cell Co-Expressed Genes Correlate With Clinical Phenotype and Microenvironments of Urothelial Cancer

doi: 10.3389/fonc.2020.553399

Figure Lengend Snippet: The results of GSEA analysis.

Article Snippet: The proteins level encoded by these genes ( PSMB10 , PSMB9 , PSMB8 , TAP1 , IRF1 , and FBXO6 ) were lower in the high clinical grade patients in the human protein atlas (HPA), which suggested the clinical phenotype correlation both in mRNA and protein levels ( ).

Techniques: