Journal: Scientific Reports

Article Title: Polyphyllin I suppresses human osteosarcoma growth by inactivation of Wnt/β-catenin pathway in vitro and in vivo

doi: 10.1038/s41598-017-07194-9

Figure Lengend Snippet: PPI suppressed osteosarcoma cells by specifically inactivating Wnt/β-catenin signaling pathway. ( A ) RT-PCR analysis of β-catenin expression level in matched human osteosarcoma tissues (tumors) and adjacent noncancerous tissues (normal) from 3 patients. ( B ) 143-B cells and ( C ) HOS cells were respectively treated with 0.8 μM PPI for indicated times, and expressions of test proteins were examined by western blotting analysis, β-actin was used as loading control, and the full-length blots were included in the supplementary information file as Figures and . 143-B cells were pretreated with 4 μM CHIR9902 (the specific GSK-3β inhibitor) for 24 h before exposed to 0.8 μM PPI for another 48 h, the combined CHIR and PPI treatments result in rescued ( D ) active β-catenin expression (the full-length blots were included in the supplementary information file as Figure ) and cell viability ( E ) compared to the PPI treatment alone in 143-B osteosarcoma cells. 143-B cells were transfected with either small interfering RNA-targeting β-catenin (si-β-catenin) or si-control for 48 h before exposed to 0.8 μM PPI for another 24 h, si-β-catenin potentiated PPI induced ( F ) down-regulation of active β-catenin expression (the full-length blots were included in the supplementary information file as Figure ), ( G ) decrease of cell viability and ( H ) inhibition of migration of 143-B osteosarcoma cells induced by PPI. * p < 0.05, ** p < 0.01, *** p < 0.001, versus the control.

Article Snippet: Briefly, 143-B cells were co-transfected with either small interfering RNA-targeting β-catenin (100 nM si-β-catenin) or 100 nM si-control for 72 h, using Lipofectamine 2000 (Invitrogen). β-catenin siRNA II (#6238) was bought from Cell Signaling Technology (Danvers, USA). si-control was designed and produced by Shanghai GenePharma Co.,Ltd (Shanghai, China).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Transfection, Small Interfering RNA, Inhibition, Migration

Journal: Oncology Letters

Article Title: Inhibitory effects of XAV939 on the proliferation of small-cell lung cancer H446 cells and Wnt/β-catenin signaling pathway in vitro

doi: 10.3892/ol.2018.8790

Figure Lengend Snippet: Reverse transcription-quantitative polymerase chain reaction assay of (A) β-catenin and (B) cyclin D1 expression. H446 cells were treated with different concentrations of XAV939 for 24 h. The expression of β-catenin and cyclin D1 compared with the blank control. The expression of β-catenin and cyclin D1 is negatively regulated by XAV939. *P<0.05 vs. control.

Article Snippet: The membranes were then incubated with the following primary antibodies: Anti-β-catenin (cat no. 6387; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-cyclin D1 (cat no. 2922; Cell Signaling Technology, Inc.) and anti-β-actin (cat no. AF0003; Beyotime Institute of Biotechnology) at a dilution of 1:1,000 at 4°C overnight.

Techniques: Real-time Polymerase Chain Reaction, Expressing

Journal: Oncology Letters

Article Title: Inhibitory effects of XAV939 on the proliferation of small-cell lung cancer H446 cells and Wnt/β-catenin signaling pathway in vitro

doi: 10.3892/ol.2018.8790

Figure Lengend Snippet: H446 cells were treated with different concentrations of XAV939 for 24 h. (A) Western blotting analysis of β-catenin and cyclin D1 expression in H446 cells compared with β-actin. The expression of (B) β-catenin and (C) cyclin D1 is inhibited by XAV939 compared with the blank control. Each bar represents the mean ± standard deviation from three independent experiments. *P<0.05.

Article Snippet: The membranes were then incubated with the following primary antibodies: Anti-β-catenin (cat no. 6387; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-cyclin D1 (cat no. 2922; Cell Signaling Technology, Inc.) and anti-β-actin (cat no. AF0003; Beyotime Institute of Biotechnology) at a dilution of 1:1,000 at 4°C overnight.

Techniques: Western Blot, Expressing, Standard Deviation

Journal: Stem Cells International

Article Title: Essentiality of CTNNB1 in Malignant Transformation of Human Embryonic Stem Cells under Long-Term Suboptimal Conditions

doi: 10.1155/2020/5823676

Figure Lengend Snippet: Suppression of multilineage differentiation capability in shCTNNB1 hESCs. (a) Immunofluorescence staining shows β -tubulin, SMA, and AFP triploblastic markers in 21-42 days differentiated hESCs (scale bar = 100 − 500 μ m). (b) Semiquantitative RT-PCR validation of mRNA expression levels of the three germ layers in 21-42 days differentiated hESCs. The error bars denote mean ± SD from three independent experiments ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, relative to WT hESCs).

Article Snippet: CTNNB1 shRNAs plasmid, pLKO.1-puro-shCTNNB1 (pLKO.1 puro shRNA beta-catenin, Plasmid #18803) were from Addgene, with empty vector (pLKO.1 puro, Plasmid, #8453) as control.

Techniques: Immunofluorescence, Staining, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Stem Cells International

Article Title: Essentiality of CTNNB1 in Malignant Transformation of Human Embryonic Stem Cells under Long-Term Suboptimal Conditions

doi: 10.1155/2020/5823676

Figure Lengend Snippet: shRNA induces the downregulation of CTNNB1 expression in hESCs. (a) Western blot analysis of CTNNB1 protein in the shCTNNB1 group, empty vector group (MOCK), and uninfected control group (WT) hESCs. (b) Immunofluorescence assay of CTNNB1 and OCT4 in shRNAs, MOCK, and WT hESCs (scale bar = 100 μ m). (c) Semiquantitative RT-PCR validation of mRNA expression levels of pluripotency genes in shCTNNB1, MOCK, and WT hESCs. Error bars denote mean ± SD from three independent experiments ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, relative to WT hESCs).

Article Snippet: CTNNB1 shRNAs plasmid, pLKO.1-puro-shCTNNB1 (pLKO.1 puro shRNA beta-catenin, Plasmid #18803) were from Addgene, with empty vector (pLKO.1 puro, Plasmid, #8453) as control.

Techniques: shRNA, Expressing, Western Blot, Plasmid Preparation, Immunofluorescence, Reverse Transcription Polymerase Chain Reaction

Journal: Stem Cells International

Article Title: Essentiality of CTNNB1 in Malignant Transformation of Human Embryonic Stem Cells under Long-Term Suboptimal Conditions

doi: 10.1155/2020/5823676

Figure Lengend Snippet: shCTNNB1 hESCs exhibit low cell proliferation and decreased telomere length as compared to WT cells. (a) Cell growth curves of shCTNNB1, MOCK, and WT hESCs. The data are represented as an average of two independent experiments ( n = 2). (b) Cell cycle analysis of shCTNNB1, MOCK, and WT hESCs. (c) Wound-healing assay. Wounds were created by manual scraping the cell clones, and images were captured as the reference point. The image of the healing state was acquired after 18 h (scale bar = 200 μ m). (d) Decreased telomerase activity in shCTNNB1 cells. Heated HSF and WT cell lysates were used as negative controls. (e) Telomere length in shCTNNB1 hESCs. Metaphase cells from the indicated groups were analyzed by the quantitative telomere-FISH assay. Strand-specific telomeric DNA probes (red) show telomere repeats; the nuclei were stained with DAPI (blue). Telomere length is represented in arbitrary units of fluorescence (a.u.) (scale bar = 10 μ m). The histograms show the frequency of different telomere lengths in CTNNB1 -silenced hESCs. The results are represented as an average of two or more independent experiments. Error bars indicate mean ± SD from three independent experiments ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01, relative to WT hESCs).

Article Snippet: CTNNB1 shRNAs plasmid, pLKO.1-puro-shCTNNB1 (pLKO.1 puro shRNA beta-catenin, Plasmid #18803) were from Addgene, with empty vector (pLKO.1 puro, Plasmid, #8453) as control.

Techniques: Cell Cycle Assay, Wound Healing Assay, Clone Assay, Activity Assay, Staining, Fluorescence