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: Stem cells (Dayton, Ohio)

Article Title: FGFR1 Signaling Stimulates Proliferation of Human Mesenchymal Stem Cells by Inhibiting the Cyclin-Dependent Kinase Inhibitors p21 Waf1 and p27 Kip1

doi: 10.1002/stem.1514

Figure Lengend Snippet: FGFR1 signaling influences the expression of S-phase kinase-associated protein two (Skp2) but not the release of Cks1. (A): Cells were transfected with vectors coexpressing a constitutively active form of FGFR1 (Fc-FGFR1), wild type (wt) FRS2, and wt Cks1. Blot depicts immunoprecipitation of Fc-R1TK (constitutively active FGFR1) 24 hours after cell synchronization and release into maintenance media alone (CTRL), or supplemented with DMSO vehicle (D) or SU5402 (SU). (B): Western blot analysis of Skp2 expression after cell synchronization and release into DMSO vehicle or SU5402 supplemented media. (C): Hypothesized pathways for FGFR1 control of cell cycle progression in hMSCs. Abbreviations: AKT, protein kinase B; Cks1, cyclin-dependent kinase subunit one; CDK, cyclin dependent kinase; FRS, fibroblast receptor substrate; FGFR1, fibroblast growth factor; ERK, extracellular signal-regulated kinase; PI3K, phosphatidylinositide 3-kinases; Rb, retinoblastoma.

Article Snippet: Rabbit anti-Cks1 (sc-6238) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, http://www.scbt.com ).

Techniques: Expressing, Transfection, Immunoprecipitation, Western Blot