eb3 gfp  (Addgene inc)

Journal: bioRxiv

Article Title: Microtubule-dependent cell polarity regulates skin-resident macrophage phagocytosis and directed cell migration

doi: 10.1101/2025.03.13.642867

Figure Lengend Snippet: A. Illustration of adult zebrafish showing overlapping scales along the trunk skin. B. Confocal images of adult trunk skin expressing reporters for Langerhans cells [Tg( mpeg1:mCherry) ] and alpha-catenin [ Gt(ctnna1-Citrine) ], which labels epidermal junctions. C. Representative image of a Langerhans cell expressing Tg(mpeg1.1:mCherry;mpeg1.1:EMTB-3xGFP) showing EMTB-3xGFP+ dendrites (yellow arrowheads), EMTB-3xGFP-dendrites (white arrows), and the presumptive microtubule organizing center (cyan arrow). D. Violin plot showing percentage of EMTB+ dendrites (n = 12 cells from N = 5 scales). E. Representative still images from time-lapse confocal microscopy of a Tg(mpeg1.1:mCherry;mpeg1.1:EB3-GFP)+ Langerhans cell showing the perinuclear focus of EB3 signal (yellow arrowhead). Yellow box in left panel denotes region magnified in the panels at right. Red and cyan arrows track individual growing microtubules. F-H. Representative still images from time-lapse confocal microscopy depicting effects of vehicle (F), nocodazole (G) , or paclitaxel (H) treatment on Tg(mpeg1.1:YFP) + Langerhans cell morphology. Yellow arrowheads in (G) indicate elongated dendrites. I-K. Violin plots showing average dendrite number (I) , average dendrite length (J) , or maximum dendrite length (K) through 80 minutes of vehicle, nocodazole, or paclitaxel treatment. n = 15 cells tracked from N = 3 scales for vehicle control, n = 27 cells tracked from N = 5 scales for nocodazole, n = 13 cells tracked from N = 6 scales for paclitaxel. L. Sholl analysis of dendrite lengths in vehicle- and nocodazole-treated conditions. n = 698 dendrites tracked in 14 cells from N = 3 scales for vehicle control and n = 476 dendrites tracked in 11 cells from N = 3 scales for nocodazole. Statistical significance in (I, J, K) was determined using Mann-Whitney U test, statistical significance in (L) was determined using a chi-squared test with the raw number of dendrites. * = p < 0.05, **** = p < 0.0001. Timestamps denote mm:ss. Scale bars, 1 mm (B), 100 μm (B’), 10 μm (B’’, C, E, F’-H’) , 20 μm (F-H).

Article Snippet: To amplify EB3-GFP, the Addgene plasmid #190164 was used as template and amplified with the primers EB3-F and EB3-R. Our previously published mpeg1.1 promoter-containing plasmid was amplified using the primers mpeg1.1_V_F’ and mpeg1.1_V_R.

Techniques: Expressing, Confocal Microscopy, Control, MANN-WHITNEY

Journal: bioRxiv

Article Title: Targeting the acetyltransferase NAT10 corrects pathologies in human frontotemporal dementia neurons and extends lifespan in an in vivo Drosophila tauopathy model

doi: 10.1101/2024.10.19.619211

Figure Lengend Snippet: A-B. Representative images of microtubule track duration (cumulative over a 150 sec interval movie), visualised by live-imaging of an EB3-GFP fusion protein, overlaid on stills from GFP-EB3 live imaging (gray) of control and FTD-MAPT-neurons (120 DIV) after 24 hours treatment with Remodelin or vehicle (DMSO). C-F. Microtubule growth, reflected in EB3 track duration, is reduced by Remodelin treatment in FTD-neurons but not in control neurons. Significance was determined using Student’s t test (ns = not significant; *p < 0.05; ****p < 0.0001); error bar represents SD; n > 15 movies from 3 independent experiments, dots represent the average of one movie).

Article Snippet: Neurons were grown to 100 DIV in individual m-Dish 35 mm dishes (Ibidi) and transfected with a plasmid encoding for GFP-EB3 (gift from Michael Davidson; Addgene plasmid # 56474).

Techniques: Imaging, Control