etc gfp β actin zip  (Addgene inc)

Journal: bioRxiv

Article Title: Bright, fluorogenic and photostable avidity probes for RNA imaging

doi: 10.1101/2021.11.02.466936

Figure Lengend Snippet: a) Confocal images of fixed MEF cells expressing GFP -β- actin-biRhoBAST 8 mRNA after incubation with smFISH probes (targeting the GFP sequence) in the presence of TMR 2 (50 nM). The overlay image shows significant colocalization of smFISH and TMR signals. Scale bar, 5 µm. b) Confocal images of live Cos7 cells co-transfected with P CMV - mAzurite-biRhoBAST 8 -MS2 24 and P UbC -NLS-stdMCP-stdGFP plasmids in the presence of 50 nM TMR 2 . The overlay image shows significant co-localization of GFP and TMR signals. Scale bar, 10 µm. c) Spinning disc confocal images of Cos7 cells expressing low levels of MCP-GFP and mAzurite-biRhoBAST 8 -MS2 24 (top), mAzurite-biRhoBAST 8 (middle), mAzurite-MS2 24 (bottom) in the presence of 500 nM TMR 2 . Single mRNA molecules were visualized with a frame rate of 10 s −1 . Scale bars, 10 µm. d) Extracted trajectories at different time points of single RNA tracks in the image shown in panel c (top), revealing synchronous movements of TMR 2 and GFP signals over time. e) Colocalization test of the cytosolic TMR 2 and GFP signals in the image shown in panel c (top), resulting in a Pearson’s R value of 0.83.

Article Snippet: To yield GFP-β-actin-Zip-biRhoBAST n , XbaI/NheI-digested biRhoBAST n cassettes were ligated into XbaI-digested and dephosphorylated eTC-GFP-β-actin-Zip (Addgene Plasmid #27123).

Techniques: Expressing, Incubation, Sequencing, Transfection

Journal: eLife

Article Title: Different translation dynamics of β- and γ-actin regulates cell migration

doi: 10.7554/eLife.68712

Figure Lengend Snippet:

Article Snippet: Constructs were generated using the eTC GFP beta-actin full-length plasmid (Addgene plasmid # 27123) and eTC GFP beta-actin ΔZip (Addgene plasmid # 27124), which were gifts from Robert Singer ( ).

Techniques: Derivative Assay, Cell Culture, Transfection, Construct, Plasmid Preparation, Transduction, Recombinant, Sequencing, Clone Assay, Software

Journal: Current Biology

Article Title: Maintenance of Miranda Localization in Drosophila Neuroblasts Involves Interaction with the Cognate mRNA

doi: 10.1016/j.cub.2017.06.016

Figure Lengend Snippet:

Article Snippet: eTC GFP beta-actin full length , [ ] , Addgene #27123.

Techniques: Recombinant, Protease Inhibitor, SYBR Green Assay, Clone Assay, Software

Journal: Scientific Reports

Article Title: Quantifying cadherin mechanotransduction machinery assembly/disassembly dynamics using fluorescence covariance analysis

doi: 10.1038/srep28822

Figure Lengend Snippet: ( a ) Top panel: Montage of live cell movie (0 hour–12 hours) showing MDCK cells expressing E-cadherin-RFP (red) and β-actin-eGFP (green) after calcium repletion. Bottom panel: +PDM maps of images from the top panel. ( b ) Left: FCI plotted every 30 minutes for E-cadherin-RFP and β-actin-eGFP after calcium repletion (n = 9). Arrow indicates the time at which FCI attains positive values. Right: FCI plotted every 5 minutes for E-cadherin-RFP and β-actin-eGFP between 2 and 2.5 hours after calcium repletion. Error bars indicate mean ± s.e.m. ( c ) Top panel: Montage of live cell movie (0 hour–12 hours) showing MDCK cells expressing E-cadherin-RFP (red) and β-actin-eGFP (green) treated with DECMA-1 antibody after calcium repletion. Bottom panel: +PDM maps of images from the top panel. ( d ) FCI values plotted every 30 minutes for E-cadherin-RFP and β-actin-eGFP after calcium repletion (n = 6). Arrow indicates the time at which FCI is positive and remains positive through the rest of the experiment. Error bars indicate mean ± s.e.m.

Article Snippet: MDCK cells stably expressing E-cadherin-RFP (a gift from Dr. James Nelson, Stanford) were transfected with the plasmid TC-eGFP-β-actin full length (Addgene ID: 27123) using Lipofectamine 2000 (Life Technologies) according to manufacturer guidelines.

Techniques: Expressing