Journal: Scientific reports

Article Title: Characterization of Three Novel H3F3A-mutated Giant Cell Tumor Cell Lines and Targeting of Their Wee1 Pathway.

doi: 10.1038/s41598-019-42611-1

Figure Lengend Snippet: Figure 3. (a) q-PCR determination of the amount of WEE1 expression in the U-GCT cell lines standardized to U-GCT1. (b) Western blot of H3.3 G34W, Wee1, H3K36me3, Rrm2 expression in the three U-GCT cell lines and A498. For the complete Western Blot see Supplementary Fig. 4. (c) Western blot of MK-1775-treated and untreated GCTB cell lines detecting H3K36me3, tyrosine 15 phosphorylated and non-phosphorylated Cdk1. For the complete Western Blot see Supplementary Fig. 5.

Article Snippet: As control cells we used the commercially available cell lines A498, Jurkat, and HEK-293 (Leibniz Institute DSMZ German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany).

Techniques: Expressing, Western Blot

Journal: Translational Andrology and Urology

Article Title: RhoJ promotes the progression of clear cell renal cell carcinoma via the TNF-α/NF-κB axis

doi: 10.21037/tau-2025-132

Figure Lengend Snippet: RhoJ knockdown inhibits the proliferation of ccRCC cells in vivo and in vitro . (A) CCK-8 assay assessing the viability of ccRCC cells. (B) Colony formation assay assessing the colony-forming ability of ccRCC cells (crystal violet staining). (C) EdU proliferation assay assessing the proliferation levels of ccRCC cells. (D) Tumor size of orthotopic xenograft models. (E,F) Cell cycle analysis of A498 (E) and 786-O (F) cells. (G) Apoptosis analysis of A498 and 786-O cells. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001; ns, not significant, P≥0.05. ccRCC, clear cell renal cell carcinoma; CCK-8, Cell Counting Kit-8; EdU, 5-ethynyl-2'-deoxyuridine; NC, negative control; RhoJ, Ras-homologous gene family member J.

Article Snippet: ccRCC cell lines (A498, 786-O) and 293T cells were cultured using Minimum Essential Medium (MEM), Roswell Park Memorial Institute-1640 (RPMI-1640) and Dulbecco’s modified Eagle medium (DMEM), respectively, under conditions of 37 °C, 5% CO 2 , and 10% fetal bovine serum (FBS; Procell, Wuhan, China).

Techniques: Knockdown, In Vivo, In Vitro, CCK-8 Assay, Colony Assay, Staining, Proliferation Assay, Cell Cycle Assay, Cell Counting, Negative Control

Journal: Translational Andrology and Urology

Article Title: RhoJ promotes the progression of clear cell renal cell carcinoma via the TNF-α/NF-κB axis

doi: 10.21037/tau-2025-132

Figure Lengend Snippet: RhoJ knockdown inhibits migration, invasion, and EMT in ccRCC cells. (A,B) Effect of RhoJ knockdown on ccRCC cell migration was assessed via wound healing assay. (A) The results were observed under an optical microscope and calculated within ImageJ. (B) Quantification of migration rate. (C,D) Effect of RhoJ knockdown on ccRCC cell migration and invasion was assessed via Transwell assay with crystal violet staining (scale bar, 20 µm; C), quantification of migration rate (D). (E) Correlation analysis between RhoJ and Bcl-2, SNAI1, VIM, and ZEB1 based on TCGA-KIRC database. (F) Western blot analyses of EMT and apoptosis-related markers in RhoJ knockdown and overexpression ccRCC cells. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001. ccRCC, clear cell renal cell carcinoma; EMT, epithelial-mesenchymal transition; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; NC, negative control; RhoJ, Ras-homologous gene family member J; SNAI1, snail family transcriptional repressor 1; TCGA-KIRC, The Cancer Genome Atlas Kidney Renal Clear Cell Carcinoma; TPM, transcript per million; VIM, Vimentin; ZEB1, zinc finger E-box binding homeobox 1.

Article Snippet: ccRCC cell lines (A498, 786-O) and 293T cells were cultured using Minimum Essential Medium (MEM), Roswell Park Memorial Institute-1640 (RPMI-1640) and Dulbecco’s modified Eagle medium (DMEM), respectively, under conditions of 37 °C, 5% CO 2 , and 10% fetal bovine serum (FBS; Procell, Wuhan, China).

Techniques: Knockdown, Migration, Wound Healing Assay, Microscopy, Transwell Assay, Staining, Western Blot, Over Expression, Negative Control, Binding Assay

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Fbxo2 inhibits cell proliferation, migration and invasion by the ubiquitin-mediated degradation of WEE1 in renal cell carcinoma

doi: 10.1007/s13402-025-01091-4

Figure Lengend Snippet: Fbxo2 overexpression inhibits the proliferation and motility of RCC cells in vitro and in vivo. (A) Western blot analysis of Fbxo2 expression in 786-O and A498 cells transfected with a Flag-Fbxo2-encoding lentivirus. (B) A CCK-8 assay was utilized to detect the effect of Fbxo2 overexpression on cell viability. (C) The proliferation of Fbxo2-overexpressing cells was measured by EdU and colony formation assays. Scale bar, 100 μm. (D) Quantitative analysis of the results of the EdU and colony formation assays. E-F : Transwell assays were performed to test the invasion capacity of RCC cells overexpressing Fbxo2. G-H : Quantification of apoptosis in RCC cells overexpressing Fbxo2 by flow cytometry. I : Sample photos of xenograft tumors in nude mice following subcutaneous injection of 786-O cells infected with an Fbxo2-overexressing lentivirus. J-K : Time course of tumor growth and tumor weights in RCC xenografts over 35 days. L : Western blot analysis of Fbxo2 expression in dissected tumors. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The human tubular cell line HK-2, along with ACHN and A498 cell lines, was acquired from Procell (Wuhan, China) and maintained in MEM (Gibco, USA) supplemented with 10% fetal bovine serum.

Techniques: Over Expression, In Vitro, In Vivo, Western Blot, Expressing, Transfection, CCK-8 Assay, Flow Cytometry, Injection, Infection

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Fbxo2 inhibits cell proliferation, migration and invasion by the ubiquitin-mediated degradation of WEE1 in renal cell carcinoma

doi: 10.1007/s13402-025-01091-4

Figure Lengend Snippet: Fbxo2 binds to and degrades WEE1. A : Coimmunoprecipitation (co-IP) was utilized to identify the interaction between endogenous Fbxo2 and WEE1 in ACHN and Caki-2 cells. B : Exogenous interaction between Fbxo2 and WEE1 in 786-O and HEK293T cells. The cells were transfected with plasmids expressing Flag-tagged Fbxo2 and Myc-tagged WEE1. C : Fbxo2 decreases endogenous WEE1 protein levels in a dose-dependent manner. HEK293T cells were transfected with increasing amounts of Flag-Fbxo2 plasmids. The cell lysates were immunoblotted after transfection for 48 h. D-E : WEE1 mRNA levels remain unaffected by Fbxo2, but WEE1 protein levels are decreased. Western blot and qRT-PCR were utilized to measure WEE1 protein and mRNA expression after overexpression of Fbxo2 in 786-O and A498 cells. F-G : Western blotting and qRT-PCR were used to assess the protein and mRNA expression levels of WEE1 after Fbxo2 was knocked down in ACHN and Caki-2 cells. H : Western blot analysis was used to measure the half-life of the WEE1 protein in HEK 293T cells with Fbxo2 knockdown. I : Quantitative plot of the abundance of WEE1 protein in (H). J : The proteasome inhibitor MG132 rescues WEE1 protein levels reduced by Fbxo2. K : HEK293T cells were transfected with Flag-Fbxo2, His-Ub, and Myc-WEE1 plasmids as indicated and treated with MG132 (10 µM) for 12 h before collection. The cell lysates and Myc immunoprecipitates were immunoblotted to detect WEE1 ubiquitination. ** p < 0.01, *** p < 0.001

Article Snippet: The human tubular cell line HK-2, along with ACHN and A498 cell lines, was acquired from Procell (Wuhan, China) and maintained in MEM (Gibco, USA) supplemented with 10% fetal bovine serum.

Techniques: Co-Immunoprecipitation Assay, Transfection, Expressing, Western Blot, Quantitative RT-PCR, Over Expression, Knockdown, Ubiquitin Proteomics