Journal: Blood

Article Title: Ruxolitinib-induced defects in DNA repair cause sensitivity to PARP inhibitors in myeloproliferative neoplasms

doi: 10.1182/blood-2017-05-784942

Figure Lengend Snippet: Sensitivity of JAK2(V617F)+, CALR(del52)+, and MPL(ex10mut)+cells to PARPi. (A) Cell lines expressing JAK2(V617F)+EpoR, CALR(del52)+MPL(wt), or MPL(W515L) were incubated with olaparib alone (1.25, 2.5, 5.0 μM) (squares) or BMN673 alone (12.5, 25.0, 50.0 nM) (squares), olaparib plus 200 μM vitamin E (circles), or olaparib plus 2.5 μM SCH51344 (triangles) for 96 hours in the presence of IL-3 plus Epo. Parental cells (diamonds) were incubated with olaparib or BMN673 only. Living cells were counted in Trypan blue. Results represent mean plus or minus SD percentage of living cells in comparison with untreated control from 3 independent experiments. (B) Western analysis of the indicated proteins in parental cells (P) and in isogenic cells expressing JAK2(V617F)+EpoR, CALR(del52)+MPL(wt), and MPL(W515L).

Article Snippet: Inhibitors/drugs The following compounds were used: JAK1/2i ruxolitinib (Selleckchem), PARPi BMN673 and PARPi olaparib (Selleckchem), mutT homolog 1 (MTH1) inhibitor SCH51344 (Tocris), ROS scavenger vitamin E (Sigma), and ribonucleoside diphosphate reductase inhibitor hydroxyurea (Selleckchem).

Techniques: Expressing, Incubation, Comparison, Control, Western Blot

Journal: The ISME Journal

Article Title: The soil microbial food web revisited: Predatory myxobacteria as keystone taxa?

doi: 10.1038/s41396-021-00958-2

Figure Lengend Snippet: Myxobacteria and potential prey organisms used in predation assays.

Article Snippet: The myxobacteria Haliangium ochraceum , Stigmatella aurantiaca , Kofleria flava , Corallococcus coralloides, and Chondromyces robustus were purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ, Table ).

Techniques: Bacteria

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: