Journal: International Journal of Molecular Sciences

Article Title: Increased H19 /miR-675 Expression in Adult T-Cell Leukemia Is Associated with a Unique Notch Signature Pathway

doi: 10.3390/ijms25105130

Figure Lengend Snippet: Adult T-cell leukemia (ATL) cells have a high notch pathway activity score. ( A ) Gene array analysis of Notch pathway activity signature genes in control, healthy peripheral blood mononuclear cells (PBMCs), two ATL lines (Jurkat, T-ALL and Nalm20, B-ALL), and three ATL lines (MT1, ATLT, and ATL25). Expression is normalized to control PBMCs and is color-coded to represent expression levels, as stated in . ( B ) Notch pathway activity score generated from array data and Qiagen pathway analysis. PBMCs served as a control, with a score of 0.0. ATL and ALL cell scores are in red and represent an elevation compared to PBMCs. p -values are indicated. ( C ) Graphs representing the Log10 normalized gene expression of Notch pathway genes of ATL cells (MT1, ATLT, and ALT25) compared to the Log10 normalized gene expression in PBMCs. Graphs are generated using Qiagen software. ( D ) Real-time PCR analysis of Notch targets in Jurkat, MT1, and ATL25. GAPDH served as a control. Results are from at least two distinct cDNA preparations, from at least two different experiments and real-time PCR runs. Cells were treated with 1 µM GSI for 48 h. Fold change is compared to cells treated with DMSO control. Significant decreases are indicated in pale green with p -values of at least > 0.05. ( E ) Venn diagram representing common GSI-sensitive targets among Jurkat (yellow), MT1 (red), and ATL25 (blue).

Article Snippet: We performed a Notch pathway activity signature array developed by Qiagen that selects 16 experimentally derived Notch signature biomarker genes, which, along with classification algorithms, acquire a quantitative analysis to determine a Notch activity score for ATL and ALL cells.

Techniques: Activity Assay, Control, Expressing, Generated, Gene Expression, Software, Real-time Polymerase Chain Reaction

Journal: Cell Death & Disease

Article Title: CircAGFG1 drives metastasis and stemness in colorectal cancer by modulating YY1/CTNNB1

doi: 10.1038/s41419-020-2707-6

Figure Lengend Snippet: a – e The proliferation, apoptosis, migration, invasion and stemness of SW480 and HCT116 cells were respectively detected through EdU assay ( a ), flow cytometry ( b ), transwell assay ( c , d ) and sphere formation assay ( e ) in sh-NC, sh-circAGFG1#1, sh-circAGFG1#1+LiCl, sh-circAGFG1#1+SC79 or sh-circAGFG1#1+Jagged1 group. f TOP/FOP-flash assay was performed to verify the activity of Wnt/β-catenin pathway in cells transfected with sh-NC or sh-circAGFG1#1. FOP-flash reporter was adopted as the control of TOP flash reporter. g The protein expressions of β-catenin, cyclin D1 and c-myc were measured by western blot in sh-NC or sh-circAGFG1#1 group. h The nuclear translocation of β-catenin in sh-NC or sh-circAGFG1#1 group was assessed by IF assay (scale bar = 20 μm). i Luciferase reporter assay was performed to verify the effect of circAFGF1 knockdown on the activity of CTNNB1 promoter. j The cellular distribution of circAGFG1 was verified by subcellular fraction assay. We repeated the experiments three times to ensure the accuracy of the experiments. ** P < 0.01.

Article Snippet: Wnt/β-catenin pathway activator LiCl, AKT activator SC79 and Notch pathway activator Jagged1 were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Migration, EdU Assay, Flow Cytometry, Transwell Assay, Tube Formation Assay, Activity Assay, Transfection, Western Blot, Translocation Assay, Luciferase, Reporter Assay