Journal: Virology Journal

Article Title: Host cell virus entry mediated by Australian bat lyssavirus G envelope glycoprotein occurs through a clathrin-mediated endocytic pathway that requires actin and Rab5

doi: 10.1186/1743-422X-11-40

Figure Lengend Snippet: Effect of dominant negative Rab GTPases on ABLV G-mediated viral entry. HEK293T cells expressing dsRed-tagged WT and DN Rab5, Rab7, and Rab11were infected with maxGFP encoding rVSV that express ABLVp G, ABLVs G, or VSV G at a MOI = 3 for 8 hrs or with rVSV that expresses EboGP at a MOI = 15 for 20 hrs and then analyzed as described in Figure . Infection of all cells was assessed. Under these conditions, the chosen MOIs yielded 40-50% virus-infected cells in controls. Results are expressed as percent virus-infected cells relative to that of WT Rab controls and represent 3 independent experiments; error bars are SEM. (A) Rab5. (B) Rab7. (C) Rab11. **, p < 0.0001; *, p < 0.005.

Article Snippet: DsRed-tagged Rab7 wild-type (WT) (plasmid #12661) and DN (T22N; plasmid #12662), DsRed-tagged Rab11 WT (plasmid #12679) and DN (S25N; plasmid #12680) [ ], and mRFP-tagged Rab5 WT (plasmid #14437) [ ] were purchased from Addgene, Cambridge, MA.

Techniques: Dominant Negative Mutation, Expressing, Infection, Virus

Journal: Nature Communications

Article Title: TGFβ signalling acts as a molecular brake of myoblast fusion

doi: 10.1038/s41467-020-20290-1

Figure Lengend Snippet: a , b , d , e DML-derived myocytes electroporated with MLC promoter driving expression of dominant negative (DN) variants of RAB11 ( b ) and RAB7 ( e ), together with membrane GFP (green) and nuclear mCherry (red). c , f , Column graph for a , b and d , e showing the population of electroporated myocytes containing the indicated number of nuclei relative to their controls (in %). g – i Functional rescue experiment where DML-derived myocytes were co-electroporated with a RFP-tagged form of TGFBR2 and an inducible (Tet-on) HA-tagged form of a constitutively active RAB7. g immunostaining against RFP showing the punctated expression of TGFBR2. h immunostaining against HA showing the diffuse expression of CA RAB7 (after Doxycyclin treatment). i Native fluorescence of H2B-BFP fusion protein, showing the nuclei within electroporated myocytes. j Merge of Fig. 5g–i. k Column graph showing the population of electroporated myocytes containing the indicated number of nuclei relative to their controls (in %) in each of the indicated conditions. Statistical analyses: DN RAB11: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\bar x$$\end{document} x ¯ : 2.57; n = 14; Ctrl: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\bar x$$\end{document} x ¯ : 2.07; n = 15; P -value <0.0001; DN RAB7: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\bar x$$\end{document} x ¯ : 1.96; n = 19; Ctrl: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\bar x$$\end{document} x ¯ : 2.46; n = 35; P -value < 0.0001; TGFBR2: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\bar x$$\end{document} x ¯ : 1.63; n = 15; Ctrl: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\bar x$$\end{document} x ¯ : 2.05; n = 23; P -value <0.0001; TGFBR2 + CA RAB7: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\bar x$$\end{document} x ¯ : 1.84; n = 27; P -value=0.0019. *** P < 0.001. ** P < 0.01. Error bars in c , f , k : SEM. Scale bars: 50 μm. Source data are provided (see ‘Data availability’).

Article Snippet: A dominant negative form of the human Rab11A (S25N) was obtained from plasmid PCMV-intron myc Rab11 S25N (a gift from Terry Hébert, Addgene plasmid #46786 ).

Techniques: Derivative Assay, Expressing, Dominant Negative Mutation, Membrane, Functional Assay, Immunostaining, Fluorescence