Journal: Cell Death & Disease

Article Title: UBE4B promotes gastric cancer proliferation and metastasis by mediating FAT4 ubiquitination and degradation

doi: 10.1038/s41419-025-07794-8

Figure Lengend Snippet: A Mass spectroscopic analysis of Up-regulated and down-regulated protein. B Volcano plot showing differentially expressed proteins ( P < 0.05, FC < 1/1.2 & FC > 1.2). C Heatmap shows a differentially expressed gene. D KEGG enrichment analysis top 20. E KEGG Level 3 shows differential protein distribution. F Western blot detected FAT4 protein in MKN45 or AGS cells after transfecting cells with three independent siRNA or UBE4B overexpression plasmid. G Western blot detected P62 and LC3II/I protein in MKN45 or AGS cells after transfecting cells with three independent siRNA or UBE4B overexpression plasmid.

Article Snippet: The antibodies used in this study were as follows: UBE4B (18148-1-AP, Proteintech), FAT4 (A17212, ABclonal), FAT4 (NBP1-78381, Novus), P62 (#7695, Cell Signaling), LC3 (#12741, Cell Signaling), ubiquitin (10201-1-AP, Proteintech), Ki67 (#9449, Cell Signaling), HA (20543-1-AP, Proteintech), MYC (390003, Zenbio), and GAPDH (HC301-01, TransGen Biotech).

Techniques: Western Blot, Over Expression, Plasmid Preparation

Journal: Cell Death & Disease

Article Title: UBE4B promotes gastric cancer proliferation and metastasis by mediating FAT4 ubiquitination and degradation

doi: 10.1038/s41419-025-07794-8

Figure Lengend Snippet: A Transmission electron microscopy observation of autophagosomes in UBE4B-knockdown MKN45 cells. B, C MKN45 cells transfected with two independent siUBE4B were treated with 20 μM cycloheximide (CHX) for 0, 2, 4, 6, 8, and 12 h, and the UBE4B and FAT4 protein levels were detected by Western blot. D Western blotting was conducted to measure FAT4 protein level after treatment with 20 μM MG132 in knockdown UBE4B and control GC cells. E, F Western blotting was conducted to measure the level of ubiquitin when knockdown or overexpression UBE4B.

Article Snippet: The antibodies used in this study were as follows: UBE4B (18148-1-AP, Proteintech), FAT4 (A17212, ABclonal), FAT4 (NBP1-78381, Novus), P62 (#7695, Cell Signaling), LC3 (#12741, Cell Signaling), ubiquitin (10201-1-AP, Proteintech), Ki67 (#9449, Cell Signaling), HA (20543-1-AP, Proteintech), MYC (390003, Zenbio), and GAPDH (HC301-01, TransGen Biotech).

Techniques: Transmission Assay, Electron Microscopy, Knockdown, Transfection, Western Blot, Control, Ubiquitin Proteomics, Over Expression

Journal: Cell Death & Disease

Article Title: UBE4B promotes gastric cancer proliferation and metastasis by mediating FAT4 ubiquitination and degradation

doi: 10.1038/s41419-025-07794-8

Figure Lengend Snippet: A Schematic representation of the docking structure of protein molecules. Blue color indicates UBE4B protein and yellow color indicates FAT4 protein. The key interacting residues are highlighted in red font. B Segment plasmid of UBE4B. C , D Endogenous UBE4B proteins were immunoprecipitated with an anti-UBE4B antibody and then analyzed by immunoblotting. Endogenous FAT4 proteins were immunoprecipitated with anti-FAT4 antibodies and then analyzed by immunoblotting. The IgG antibody was used as the control. E HEK-293T cells infected endogenous overexpression of FAT4 lentivirus and HA-UBE4B plasmid were subject to immunoprecipitation with anti-UBE4B and anti-FAT4 antibodies. F HEK-293T cells infected endogenous overexpression of FAT4 lentivirus were transfected with HA-UBE4B or several UBE4B mutants. A coimmunoprecipitation assay was performed to detect the interaction between UBE4B and FAT4 protein. G HEK-293T cells infected with endogenous overexpression of FAT4 lentivirus were transfected with HA-UBE4B or wild-type Ub, Ub-K6R, K11R, K27R, K29R, K33R, K48R, and K63R plasmid. Cell lysates were assayed for ubiquitination, and the ubiquitination level of FAT4 was detected. H HEK-293T cells infected with endogenous overexpression of FAT4 lentivirus were transfected with HA-UBE4B or wild-type Ub, Ub-K6R, K33R and K6R/K33R plasmid. Cell lysates were assayed for ubiquitination, and the ubiquitination level of FAT4 was detected.

Article Snippet: The antibodies used in this study were as follows: UBE4B (18148-1-AP, Proteintech), FAT4 (A17212, ABclonal), FAT4 (NBP1-78381, Novus), P62 (#7695, Cell Signaling), LC3 (#12741, Cell Signaling), ubiquitin (10201-1-AP, Proteintech), Ki67 (#9449, Cell Signaling), HA (20543-1-AP, Proteintech), MYC (390003, Zenbio), and GAPDH (HC301-01, TransGen Biotech).

Techniques: Plasmid Preparation, Immunoprecipitation, Western Blot, Control, Infection, Over Expression, Transfection, Co-Immunoprecipitation Assay, Ubiquitin Proteomics

Journal: Cell Death & Disease

Article Title: UBE4B promotes gastric cancer proliferation and metastasis by mediating FAT4 ubiquitination and degradation

doi: 10.1038/s41419-025-07794-8

Figure Lengend Snippet: A MKN45 cells with/without shUBE4B were injected in nude mice. Representative images of xenograft tumors from mice bearing MKN45-shNC and MKN45-shUBE4B cells. B Tumor weights were calculated for each mouse in both groups ( n = 6, Student’s t test). C Tumor volume was measured every 5 days ( n = 6, Student’s t test). D IHC scores were analyzed for the expression levels of UBE4B, FAT4, and Ki67 in both groups (NC vs UBE4B-OE, n = 15, Student’s t test). E , F UBE4B, FAT4, and Ki67 expression in the tumor of nude mice were detected by IHC. Bar length: 50 μm. G MKN45 cells with/without shUBE4B or with/without FAT4 were injected in nude mice. Representative images of xenograft tumors from mice bearing MKN45-shNC+shNC, MKN45-shUBE4B+shNC, and MKN45-shUBE4B+shFAT4 cells. H Tumor weights were calculated for each mouse in the three groups ( n = 10, One-way ANOVA). I Tumor volume was measured every 3 days ( n = 10, One-way ANOVA). J IHC scores were analyzed for the expression levels of UBE4B, FAT4, and Ki67 in the three groups (NC + NC vs KD-UBE4B + NC vs KD-UBE4B + KD-FAT4, n = 15, One-way ANOVA).

Article Snippet: The antibodies used in this study were as follows: UBE4B (18148-1-AP, Proteintech), FAT4 (A17212, ABclonal), FAT4 (NBP1-78381, Novus), P62 (#7695, Cell Signaling), LC3 (#12741, Cell Signaling), ubiquitin (10201-1-AP, Proteintech), Ki67 (#9449, Cell Signaling), HA (20543-1-AP, Proteintech), MYC (390003, Zenbio), and GAPDH (HC301-01, TransGen Biotech).

Techniques: Injection, Expressing

Journal: Cell Death & Disease

Article Title: UBE4B promotes gastric cancer proliferation and metastasis by mediating FAT4 ubiquitination and degradation

doi: 10.1038/s41419-025-07794-8

Figure Lengend Snippet: A Representative image of IHC staining of tumor microarrays from GC patients. Bar length: 50 μm. B IHC scores of UBE4B level between GC tissue and adjacent normal tissue. C IHC scores of FAT4 level between GC tissue and adjacent normal tissue. D The Kaplan-Meier plot of overall survival by the expression of UBE4B in tumor microarrays related information ( n = 66 (high-expression) vs 28 (low-expression)). E The Kaplan-Meier plot of overall survival by the expression of FAT4 in tumor microarrays related information ( n = 46 (high-expression) vs 48 (low-expression)).

Article Snippet: The antibodies used in this study were as follows: UBE4B (18148-1-AP, Proteintech), FAT4 (A17212, ABclonal), FAT4 (NBP1-78381, Novus), P62 (#7695, Cell Signaling), LC3 (#12741, Cell Signaling), ubiquitin (10201-1-AP, Proteintech), Ki67 (#9449, Cell Signaling), HA (20543-1-AP, Proteintech), MYC (390003, Zenbio), and GAPDH (HC301-01, TransGen Biotech).

Techniques: Immunohistochemistry, Expressing

Journal: Journal of Gastric Cancer

Article Title: Loss of FAT Atypical Cadherin 4 Expression Is Associated with High Pathologic T Stage in Radically Resected Gastric Cancer

doi: 10.5230/jgc.2015.15.1.39

Figure Lengend Snippet: Immunohistochemical staining of FAT4 in normal gastric mucosa and gastric adenocarcinoma (×200). FAT4 expression was well preserved in normal gastric mucosa (A), while tumors showed variable expressions of FAT4. Strong and diffused FAT4 expression was seen in intestinal-type gastric adenocarcinoma (B), while focal FAT4 expression was observed in intestinal- (C) and diffuse-type (D) gastric adenocarcinoma. A total loss of the expression of FAT4 was also observed in intestinal- (E) and diffuse-type (F) gastric adenocarcinoma.

Article Snippet: Sections were washed, and stained with a rabbit polyclonal antibody against FAT4 (NBP1-78381; Novus biological, Littleton, CO, USA) diluted 1:50.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Journal of Gastric Cancer

Article Title: Loss of FAT Atypical Cadherin 4 Expression Is Associated with High Pathologic T Stage in Radically Resected Gastric Cancer

doi: 10.5230/jgc.2015.15.1.39

Figure Lengend Snippet: Comparison of the H-scores of FAT4 immunohistochemical staining. (A) The H-scores for FAT4 in normal gastric mucosa (mean H-score, 148.1±67.4) and gastric cancer (mean H-score, 60.0±63.1). (B) The H-scores of FAT4 according to the pathologic T stage. (C) The H-scores of FAT4 according to the American Joint Committee on Cancer (AJCC) cancer staging system. * P<0.05.

Article Snippet: Sections were washed, and stained with a rabbit polyclonal antibody against FAT4 (NBP1-78381; Novus biological, Littleton, CO, USA) diluted 1:50.

Techniques: Comparison, Immunohistochemical staining, Staining

Journal: Journal of Gastric Cancer

Article Title: Loss of FAT Atypical Cadherin 4 Expression Is Associated with High Pathologic T Stage in Radically Resected Gastric Cancer

doi: 10.5230/jgc.2015.15.1.39

Figure Lengend Snippet: Expression of FAT4 in relation to clinicopathological parameters in GCs

Article Snippet: Sections were washed, and stained with a rabbit polyclonal antibody against FAT4 (NBP1-78381; Novus biological, Littleton, CO, USA) diluted 1:50.

Techniques: Expressing

Journal: Journal of Gastric Cancer

Article Title: Loss of FAT Atypical Cadherin 4 Expression Is Associated with High Pathologic T Stage in Radically Resected Gastric Cancer

doi: 10.5230/jgc.2015.15.1.39

Figure Lengend Snippet: Disease-free survival (DFS) and overall survival (OS) in relation to FAT4 expression in gastric cancer. (A) The loss of FAT4 expression was significantly associated with poor DFS (5-year survival rate; loss vs. no loss, 68.7% vs. 86.9%, P=0.017). (B) The FAT4-negative group was associated with poor OS, but statistical significance was borderline (5-year survival rate; loss vs. no loss, 57.6% vs. 70.9%, P=0.083). * P<0.05.

Article Snippet: Sections were washed, and stained with a rabbit polyclonal antibody against FAT4 (NBP1-78381; Novus biological, Littleton, CO, USA) diluted 1:50.

Techniques: Expressing