Journal: Oncology Letters

Article Title: Tumor-suppressive microRNA-452 inhibits migration and invasion of breast cancer cells by directly targeting RAB11A

doi: 10.3892/ol.2017.6426

Figure Lengend Snippet: miR-452 mimic sequence alongside RAB11A-3′-UTR-WT and RAB11A-3′-UTR-Mut mRNA sequences, including the miR-452-binding site of the WT sequence, and the equivalent area from the Mut sequence. miR-452, microRNA-452; 3′-UTR, 3′-untranslated region; WT, wild type; Mut, mutant.

Article Snippet: Luciferase reporter assay Plasmids containing the wild-type RAB11A 3′-UTR (PmirGLO-RAB11A-3′-UTR-WT) or a mutant RAB11A 3′-UTR (PmirGLO-RAB11A-3′-UTR-Mut; ) were obtained from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Sequencing, Binding Assay, Mutagenesis

Journal: Oncology Letters

Article Title: Tumor-suppressive microRNA-452 inhibits migration and invasion of breast cancer cells by directly targeting RAB11A

doi: 10.3892/ol.2017.6426

Figure Lengend Snippet: miR-452 directly targets RAB11A in breast cancer cells. The relative RAB11A (A) mRNA and (B) protein expression levels were significantly reduced in the miR-452 mimic-transfected breast cancer cells, compared with miR-NC-transfected cells. (C) A luciferase reporter assay was performed on HEK293T cells co-transfected with an miR-452 mimic or miR-NC, and PmirGLO-RAB11A-3′-UTR-WT or PmirGLO-RAB11A-3′-UTR-Mut, revealing that miR-452 decreased activity only in cells transfected with the WT RAB11A 3′-UTR. Data are presented as the mean ± standard deviation. *P<0.05 vs. corresponding miR-NC group. miR-452, microRNA-452; miR-NC, microRNA negative control; 3′-UTR, 3′-untranslated region; WT, wild type; Mut, mutant.

Article Snippet: Luciferase reporter assay Plasmids containing the wild-type RAB11A 3′-UTR (PmirGLO-RAB11A-3′-UTR-WT) or a mutant RAB11A 3′-UTR (PmirGLO-RAB11A-3′-UTR-Mut; ) were obtained from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Expressing, Transfection, Luciferase, Reporter Assay, Activity Assay, Standard Deviation, Negative Control, Mutagenesis

Journal: Oncology Letters

Article Title: Tumor-suppressive microRNA-452 inhibits migration and invasion of breast cancer cells by directly targeting RAB11A

doi: 10.3892/ol.2017.6426

Figure Lengend Snippet: Inhibition of RAB11A inhibits migration and invasion of MCF-7 and MDA-MB-231 breast cancer cells. (A) Compared with siR-NC, RAB11A siRNA decreased RAB11A protein expression in breast cancer cells. (B) Transwell migration and (C) Transwell invasion assays were performed, revealing that RAB11A knockdown decreased breast cancer cell migration and invasion capacities. Data are presented as the mean ± standard deviation. *P<0.05 vs. corresponding siR-NC group. siRNA, small interfering RNA; siR-NC, negative control siRNA.

Article Snippet: Luciferase reporter assay Plasmids containing the wild-type RAB11A 3′-UTR (PmirGLO-RAB11A-3′-UTR-WT) or a mutant RAB11A 3′-UTR (PmirGLO-RAB11A-3′-UTR-Mut; ) were obtained from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Inhibition, Migration, Expressing, Knockdown, Standard Deviation, Small Interfering RNA, Negative Control

Journal: The EMBO Journal

Article Title: TDP‐43 loss of function inhibits endosomal trafficking and alters trophic signaling in neurons

doi: 10.15252/embj.201694221

Figure Lengend Snippet: Primary hippocampal neurons (DIV6+3) were transfected with either shTDP or shCtrl together with GFP‐tagged organelle markers. A–C Mito‐GFP visualizes trafficking of mitochondria. At least four dendrite segments per neuron were live‐imaged with 2 Hz for 2.5 min. Quantitative analysis of relative vesicle movement (B) and vesicle number (C) from kymographs. D–F GFP‐RAB5 visualizes early endosomes. At least four dendrite segments per neuron were live‐imaged with 2 Hz for 2.5 min. Quantitative analysis of relative vesicle movement (E) and vesicle number (F) from kymographs. G–I GFP‐RAB4 visualizes fast recycling endosomes. At least four dendrite segments per neuron were live‐imaged with 2 Hz for 2.5 min. Quantitative analysis of relative vesicle movement (H) and vesicle number (I) from kymographs. Data information: Mean ± s.e.m., n = 3, unpaired, two‐tailed t ‐test: no significant differences were detected.

Article Snippet: Human RAB4 (Addgene #54943, M. Davidson unpublished), RAB5 (Lang et al , ), RAB7 and RAB11a wild type and S25N (Addgene #12605, 12674, 12678, Richard Pagano (Choudhury et al , )) were expressed from pEGFP‐C1 vector.

Techniques: Transfection, Two Tailed Test