Journal: Scientific reports

Article Title: Tripartite motif 31 drives gastric cancer cell proliferation and invasion through activating the Wnt/β-catenin pathway by regulating Axin1 protein stability.

doi: 10.1038/s41598-023-47139-z

Figure Lengend Snippet: Figure 4. TRIM31 activates the Wnt/β-catenin pathway by enhancing Axin1 ubiquitination and degradation. (A) Relative protein expression levels of core Wnt/β-catenin pathway components and TRIM31 upon transfection in HGC-27 cells, as determined by western blotting assays. (B) The cytoplasmic/nuclear fractionation of β-catenin protein under TRIM31-knockdown and overexpression. GAPDH served as the cytoplasmic marker, while Histone H3 functioned as the nuclear marker. (C,D) The mRNA expression of TRIM31 and core components of the Wnt/β-catenin signaling pathway detected via qRT-PCR assays using modified HGC-27 cells after transfection. (E) Axin1 protein expression in modified HGC-27 cells with 100 μg/ ml CHX at the indicated time points. (F) Line graph of Axin1 protein expression standardized with GAPDH control group at corresponding time points. (G) The in vivo ubiquitination assays were performed to detect the Axin1 protein expression upon TRIM31 knockdown. Indicated antibodies were utilized for detecting the input and binding proteins using WB analysis. (H) Interaction between TRIM31 and Axin1 in HGC-27 cells via co-IP assays (ns no significance, *P < 0.05 and **P < 0.01).

Article Snippet: All primary antibodies used in this study were as follows: TRIM31 (1:2000, Abcam, ab67785), β-catenin (1:2000, BD, 610,153), Histone H3 (1:1000, CST, 4499), c-Myc (1:2000, Affinity, AF0358), GSK-3β (1:1000, CST, 9315), Axin1 (1:1000, Fitzgerald, 70R-51344), Axin2 (1:2000, Prosci, 6163), cyclinD1 (1:5000, Proteintech, 60186-1-Ig), β-actin (1:5000, Proteintech, 20536-1-AP) and GAPDH (1:5000, Proteintech, 60004-1-Ig). β-actin and GAPDH were used as negative controls.

Techniques: Ubiquitin Proteomics, Expressing, Transfection, Western Blot, Fractionation, Knockdown, Over Expression, Marker, Quantitative RT-PCR, Modification, Control, In Vivo, Binding Assay, Co-Immunoprecipitation Assay

Journal: Scientific reports

Article Title: Tripartite motif 31 drives gastric cancer cell proliferation and invasion through activating the Wnt/β-catenin pathway by regulating Axin1 protein stability.

doi: 10.1038/s41598-023-47139-z

Figure Lengend Snippet: Figure 6. TRIM31 was negatively correlated with Axin1 protein expression in GC tissues. (A) Analysis of TRIM31 and Axin1 expression in 8 freshly collected GC tissues using western blotting assays. (B) Expression analysis of TRIM31 and Axin1 protein levels in 8 GC patients by the Pearson correlation test.

Article Snippet: All primary antibodies used in this study were as follows: TRIM31 (1:2000, Abcam, ab67785), β-catenin (1:2000, BD, 610,153), Histone H3 (1:1000, CST, 4499), c-Myc (1:2000, Affinity, AF0358), GSK-3β (1:1000, CST, 9315), Axin1 (1:1000, Fitzgerald, 70R-51344), Axin2 (1:2000, Prosci, 6163), cyclinD1 (1:5000, Proteintech, 60186-1-Ig), β-actin (1:5000, Proteintech, 20536-1-AP) and GAPDH (1:5000, Proteintech, 60004-1-Ig). β-actin and GAPDH were used as negative controls.

Techniques: Expressing, Western Blot

Journal: Scientific reports

Article Title: Tripartite motif 31 drives gastric cancer cell proliferation and invasion through activating the Wnt/β-catenin pathway by regulating Axin1 protein stability.

doi: 10.1038/s41598-023-47139-z

Figure Lengend Snippet: Figure 5. Axin1 silencing rescues the proliferation and invasion of TRIM31 knockdown in GC cells. (A) Relative protein expression levels of TRIM31, Axin1, and main Wnt pathway components were analyzed through WB assays upon transfection. (B,C) Representative images and relative quantification of HGC-27 and BGC823 cells from colony-formation experiments. (D,E) Representative images of GC cells from transwell invasion analysis (magnification, × 200). Bar graphs of the invasion data. (F,G) Wound healing assays revealed the migratory capabilities of modified GC cells. The relative quantification was shown in the bar graph (One- way ANOVA: *P < 0.05 and **P < 0.01).

Article Snippet: All primary antibodies used in this study were as follows: TRIM31 (1:2000, Abcam, ab67785), β-catenin (1:2000, BD, 610,153), Histone H3 (1:1000, CST, 4499), c-Myc (1:2000, Affinity, AF0358), GSK-3β (1:1000, CST, 9315), Axin1 (1:1000, Fitzgerald, 70R-51344), Axin2 (1:2000, Prosci, 6163), cyclinD1 (1:5000, Proteintech, 60186-1-Ig), β-actin (1:5000, Proteintech, 20536-1-AP) and GAPDH (1:5000, Proteintech, 60004-1-Ig). β-actin and GAPDH were used as negative controls.

Techniques: Knockdown, Expressing, Transfection, Quantitative Proteomics, Modification

Journal: Scientific reports

Article Title: Tripartite motif 31 drives gastric cancer cell proliferation and invasion through activating the Wnt/β-catenin pathway by regulating Axin1 protein stability.

doi: 10.1038/s41598-023-47139-z

Figure Lengend Snippet: Figure 7. A model of TRIM31 regulation of Axin1-Wnt/β-catenin pathway in GC.

Article Snippet: All primary antibodies used in this study were as follows: TRIM31 (1:2000, Abcam, ab67785), β-catenin (1:2000, BD, 610,153), Histone H3 (1:1000, CST, 4499), c-Myc (1:2000, Affinity, AF0358), GSK-3β (1:1000, CST, 9315), Axin1 (1:1000, Fitzgerald, 70R-51344), Axin2 (1:2000, Prosci, 6163), cyclinD1 (1:5000, Proteintech, 60186-1-Ig), β-actin (1:5000, Proteintech, 20536-1-AP) and GAPDH (1:5000, Proteintech, 60004-1-Ig). β-actin and GAPDH were used as negative controls.

Techniques:

Journal: Microorganisms

Article Title: Myxobacterial Predation: A Standardised Lawn Predation Assay Highlights Strains with Unusually Efficient Predatory Activity

doi: 10.3390/microorganisms11020398

Figure Lengend Snippet: Myxobacteria strains used. DSMZ refers to strains obtained from the German Collection of Microorganisms and Cell Cultures. Myxococcus llanfairPGensis is an abbreviation of Myxococcus llanfairpwllgwyngyllgogerychwyrndrobwllllantysiliogogogochensis .

Article Snippet: Myxococcia , Corallococcus , Corallococcus coralloides DSM 2259 , DSMZ , GCA_000255295.

Techniques: