Journal: Brain and Behavior

Article Title: E2F transcription factor 1 elevates cyclin D1 expression by suppressing transcription of microRNA‐107 to augment progression of glioma

doi: 10.1002/brb3.2399

Figure Lengend Snippet: E2F1 mediates the Wnt/β‐catenin signaling pathway. (a) Nuclear translocation of β‐catenin in A172 and T98G cells examined by immunofluorescence staining; (b) TOP/FOP activity in A172 and T98G cells determined by the TOP/FOP flash assay; (c) Protein levels of Wnt10B and β‐catenin in A172 and T98G cells detected by western blot analysis. Data were collected from three independent experiments and presented as mean ± SD. Differences were compared by one‐way ANOVA (b) or two‐way ANOVA (c), * p < .05 versus miR‐107 mimic +oe‐NC

Article Snippet: Cells in each group were cultured in 48‐well plates at a density of 2 × 10 4 cells per well for 24 h. Each well was loaded with 0.2 μg TOP/FOP flash and 1 ng Renilla (pRLTK) luciferase‐encoding plasmid (Promega) according to the instructions of a Lipofectamine 3000 kit (Thermo Fisher Scientific Inc., Waltham, MA, USA).

Techniques: Translocation Assay, Immunofluorescence, Staining, Activity Assay, Western Blot

Journal: Biology Open

Article Title: Rab7 is required for mesoderm patterning and gastrulation in Xenopus

doi: 10.1242/bio.056887

Figure Lengend Snippet: Rab7 is required for canonical Wnt pathway activation. (A) Wild-type dorsal expression of chd in comparison to specimen with (B) radial injected wnt8a mRNA showing an extended chd expression domain around the blastopore. (C) Co-injection of rab7 TBMO restricted additional chd expression to normal wild-type state. (D) Quantification of results in A-C. (E) Untreated control specimen, (F) in comparison to induced double axis (white arrowhead) after ventral wnt3a mRNA injection, (G) parallel injection of rab7 TBMO inhibited secondary axis formation (black arrowhead). (H) Quantification of results in E-G. (I) Luciferase-based Wnt reporter assay analysis at stage 10.5 illustrated that wnt8a induced reporter activity was blocked by rab7 loss of function in animal caps. (J,N) Wild-type expression of myod1 and myf5 in stage 11 control embryos. (K,O) Absent dorsal myod1 expression and lost domains upon rab7 loss of function (black arrowheads). (L,P) Treatment with LiCl partially rescues dorsal expression of myod1 and myf5 in rab7 morphant embryos (white arrowheads). Please note that LiCl-induced ectopic expression in the axial mesoderm was not inhibited by the rab7 TBMO. (M,Q) Quantification of results in J-L and N-P, respectively. Scale bars: 250 µm.

Article Snippet: For Luciferase-based Wnt assay (Promega Dual-Luciferase ® Reporter Assay System), embryos were injected into two animal or ventral blastomeres at the four-cell stage (80 pg BAR reporter DNA plus 40 pg Renilla DNA).

Techniques: Activation Assay, Expressing, Injection, Luciferase, Reporter Assay, Activity Assay