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: Biochimica et biophysica acta

Article Title: SNARE-mediated membrane traffic is required for focal adhesion kinase signaling and Src-regulated focal adhesion turnover.

doi: 10.1016/j.bbamcr.2010.09.008

Figure Lengend Snippet: Fig. 5. Inhibition of SNARE-mediated membrane traffic inhibits Src trafficking from a Rab11 compartment. CHO-E329Q cells were mock treated (Wild-type NSF; A–C, G–I) or induced with doxycycline (E329Q-NSF; D–F, J–L) for 9 h and serum-starved for the last hour, before being lifted, and plated on FN for the indicated times. Cells were fixed in ice-cold 0.1% Triton X-100, 2% paraformaldehyde and stained with anti-Src (red; A, D, G, and J) and anti-Rab11 (green; B, E, H, and K). Arrowheads point to Src-Rab11-containing compartment in WT-NSF cells (I) and E329Q-NSF cells (L). Note the increased amount of Src in Rab11-containing compartment in L. Scale bar=10 μm. (M) Cells from the above experiments, as well as untreated cells, and cells where membrane traffic was inhibited with truncated SNAP23 (SN23CΔ9) and dominant-negative Rab11 (Rab11S25N) were visually scored as having primarily perinuclear Src staining (light grey bars), as seen in panel A, or peripheral Src staining (dark grey bars), as seen in panel G. Results are the means±SEM of at least 3 separate experiments in which at least 50 cells were scored.

Article Snippet: Antibodies were obtained against NSF from Stressgen Biotechnologies (Victoria, BC), Rab11, phosphoTyr397-FAK and mouse anti-FAK (F15020) from BD Biosciences (Mississauga, ON), rabbit anti-FAK from Abcam (ab40794; Cambridge, MA), phosphoTyr576-FAK from Novus Biologicals (Littleton, CO), phosphoTyr925-FAK and phosphoSer473Akt from Cell Signaling Technology, Inc. (Boston, MA), phosphoTyr418Src, phosphoTyr861-FAK and Src from Applied Biological Materials, Inc. (Richmond, BC), α-tubulin from Invitrogen, Co. (A11126; Mississauga, ON), and Rac from Cytoskeleton, Inc. (Denver, CO).

Techniques: Inhibition, Membrane, Staining, Dominant Negative Mutation