Journal: Cell Death & Disease

Article Title: Angiomotin-p130 inhibits β-catenin stability by competing with Axin for binding to tankyrase in breast cancer

doi: 10.1038/s41419-019-1427-2

Figure Lengend Snippet: a A visualized heatmap of differentially expressed genes in MCF7KD cells compared to MCF7 control cells. Red color indicates upregulation and green indicates downregulation (detailed data are uploaded as Supplementary Table ). b Functionally associated pathways were identified by Ingenuity Pathway Analysis. Enriched pathways related to breast carcinogenesis were sorted by Fisher's exact P values. c An interactive network analysis of the key molecular β-catenin from the first ranked WNT pathway. d Expression validation of the genes from the interactive network by quantitative PCR. The means ± SD of relative fold changes from triplicate experiments were plotted. GAPDH was used as the control. P values were calculated by paired Student’s t test

Article Snippet: Antibodies against β-catenin (#8084P), p-β-catenin (#9561T), YAP (#14074S), TAZ (#4883), vimentin (#5741P), E-cadherin (#3195P), OCT-4 (#2750), Nanog (#4903P), SOX2 (#3579P), C-myc (#5605), Cyclin D (#2978), LEF1 (#2230P), TCF4 (#2565), MET (#8198), Gsk3β (#12456T), and β-trcp (#4394S) were from Cell Signaling Technology.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Cell Death & Disease

Article Title: Angiomotin-p130 inhibits β-catenin stability by competing with Axin for binding to tankyrase in breast cancer

doi: 10.1038/s41419-019-1427-2

Figure Lengend Snippet: a Protein levels of total β-catenin, cytoplasmic β-catenin, and nuclear β-catenin as determined by western blotting. GAPDH was used as the loading control for the total and cytoplasmic protein. Lamin A was used as the loading control for nuclear protein. b β-Catenin-driven transcription activity was determined by TOP/FOP luciferase reporter assays. Normalization was based on internal Renilla luciferase actvity. The final reporter activity was measured as TOP/FOP ratio and was expressed as the mean ± SD of three independent experiments. c Protein levels of WNT downstream targets were determined by western blotting. d β-Catenin expression was determined by immunohistochemistry staining in xenograft tissues. Scale bar = 200 µm. e Co-localization of Amot-p130 (red) and β-catenin (green) in cell–cell contacts was indicated by immunofluorescence confocal microscopy. DAPI was used for nuclear staining (blue). f The interaction between Amot-p130 and β-catenin was evaluated in MCF7 cells by co-immunoprecipitation assay. IgG was used as the negative control. *** P < 0.001. DAPI, 4′,6-diamidino-2-phenylin

Article Snippet: Antibodies against β-catenin (#8084P), p-β-catenin (#9561T), YAP (#14074S), TAZ (#4883), vimentin (#5741P), E-cadherin (#3195P), OCT-4 (#2750), Nanog (#4903P), SOX2 (#3579P), C-myc (#5605), Cyclin D (#2978), LEF1 (#2230P), TCF4 (#2565), MET (#8198), Gsk3β (#12456T), and β-trcp (#4394S) were from Cell Signaling Technology.

Techniques: Western Blot, Activity Assay, Luciferase, Expressing, Immunohistochemistry, Staining, Immunofluorescence, Confocal Microscopy, Co-Immunoprecipitation Assay, Negative Control

Journal: Cell Death & Disease

Article Title: Angiomotin-p130 inhibits β-catenin stability by competing with Axin for binding to tankyrase in breast cancer

doi: 10.1038/s41419-019-1427-2

Figure Lengend Snippet: a β-Catenin levels in cells treated with CHX (200 µg/ml) for the indicated time, with or without MG132 (20 µM) for 2 h, were determined using western blotting (left). GAPDH was used as the loading control. The curve showed the relative trend of β-catenin changes (right). b β-Catenin levels in cells treated with XAV939 10 µg/ml for 24 h were determined using western blotting (left). The quantitation of β-catenin was expressed as the mean ± SD of three independent experiments (right). c β-catenin levels in cells treated with SKL2001 30 µM for 24 h were determined using western blotting (left). The quantitation of β-catenin was expressed as the mean ± SD of three independent experiments (right). d Protein levels of the total, cytoplasmic, and nuclear β-catenin in cells treated with XAV939 or SKL2001, alone or in combination with MG132, were determeined using western blotting. GAPDH was used as the loading control for the total and cytoplasmic protein. Lamin A was used as the loading control for nuclear protein. e MCF7 cell proliferation under XAV939 or SKL2001 treatment was determined by MTT assay. f Relative quantitation of Amot-p130 and Axin pulled down by TNKS in co-immunoprecipitation assay. IgG pull-down was used as the negative control. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, no significance. MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; TNKS, tankyrase

Article Snippet: Antibodies against β-catenin (#8084P), p-β-catenin (#9561T), YAP (#14074S), TAZ (#4883), vimentin (#5741P), E-cadherin (#3195P), OCT-4 (#2750), Nanog (#4903P), SOX2 (#3579P), C-myc (#5605), Cyclin D (#2978), LEF1 (#2230P), TCF4 (#2565), MET (#8198), Gsk3β (#12456T), and β-trcp (#4394S) were from Cell Signaling Technology.

Techniques: Western Blot, Quantitation Assay, MTT Assay, Co-Immunoprecipitation Assay, Negative Control

Journal: Cell Death & Disease

Article Title: Angiomotin-p130 inhibits β-catenin stability by competing with Axin for binding to tankyrase in breast cancer

doi: 10.1038/s41419-019-1427-2

Figure Lengend Snippet: Axin/TNKS binding contributes to the nuclear translocation of β-catenin and the further activation of the WNT pathway in Amot-p130 negative cells (left). Amot-p130/TNKS binding allows Axin to form β-catenin-destruction complexes, leading to the phosphorylation and ubiquitination-mediated degradation of β-catenin, thereby inactivating WNT signaling in Amot-p130 positive cells (right). TNKS, tankyrase

Article Snippet: Antibodies against β-catenin (#8084P), p-β-catenin (#9561T), YAP (#14074S), TAZ (#4883), vimentin (#5741P), E-cadherin (#3195P), OCT-4 (#2750), Nanog (#4903P), SOX2 (#3579P), C-myc (#5605), Cyclin D (#2978), LEF1 (#2230P), TCF4 (#2565), MET (#8198), Gsk3β (#12456T), and β-trcp (#4394S) were from Cell Signaling Technology.

Techniques: Binding Assay, Translocation Assay, Activation Assay

Journal: Molecular medicine reports

Article Title: Naringin ameliorates memory deficits and exerts neuroprotective effects in a mouse model of Alzheimer's disease by regulating multiple metabolic pathways.

doi: 10.3892/mmr.2021.11971

Figure Lengend Snippet: Figure 8. Naringin exerts neuroprotective effects by regulating the glutamate receptor system and apoptosis. (A) Protein levels of NMDAR1, GluR2 and CAMKII. (B) Protein levels of Bad, Bcl‑2 and cleaved‑caspase‑3. **P<0.01 vs. sham group; #P<0.05 and ##P<0.01 vs. model group; &P<0.05, &&P<0.01 vs. naringin group. NMDAR1, glutamate receptor subunit 1; GluR2, glutamate receptor 2; CAMKII, calcium/calmodulin‑dependent protein kinase type II.

Article Snippet: After blocking with 5% non‐fat dried milk for 2.5 h at 37 ̊C, the PVDF membrane was incubated with primary antibodies against: APP (1:1,000; cat. no. bs‐12503R; BIOSS), BACE1 (1:1,000; cat. no. 5606T; Cell Signaling Technology, Inc.), CDK5 (1:1,000; cat. no. bs‐10258Rm; BIOSS), p‐Tau396 (1:1,000; cat. no. bs‐3446R; BIOSS), glutamate receptor subunit 1 (NMDAR1) (1:1,000; cat. no. bs‐23343R; BIOSS), glutamate receptor 2 (GluR2; 1:1,000; cat. no. bs‐13385R; BIOSS), calcium/calmod‐ ulin‐dependent protein kinase type II (CAMKII; 1:1,000; Boster), Bad (1:500; cat. no. A00183; Boster), caspase‐3 (1:500; cat. no. bs‐0081R; BIOSS), Bcl‐2 (1:500; cat. no. bs‐0032R; BIOSS), ERβ (1:1,000; cat. no. kl437Hu22; KALANG; https://www.biomart.cn/infosupply/31407572.htm), p‐P38 (1:1,000; cat. no. bs‐2210R; BIOSS) and β‐actin (1:1,000; cat. no. bs‐0061R; BIOSS) overnight.

Techniques: