Journal: Journal of Cell Communication and Signaling
Article Title: Myosin-X is essential to the intercellular spread of HIV-1 Nef through tunneling nanotubes
doi: 10.1007/s12079-018-0493-z
Figure Lengend Snippet: Western blot analyses of Myo10 expression in (a) mouse neuronal CAD cells and (b) human epithelial HeLa cells transfected with GFP-vector (control) and Nef-GFP are shown. Calnexin was used as loading controls. Blots are representative of 3 independent experiments, P value = 0.005 (*). In both cell types, Myo10 expression increases upon Nef-GFP expression. Human MDM were infected with reverse transcriptase-normalized VSV-G pseudotyped virus stocks (WT = pNL4–3 clone) or (DelNef = pNL4–3delNef clone) at 5, 10, and 20 cpm of reverse transcriptase activity/cell. (c) A representative Western blot of the WT or DelNef infected cells from 3 independent infections is shown along with (d) Densitometry representation of the relative expression levels of Myo10 normalized to the amount of tubulin (loading control), with a P value <0.01 (**) or < 0.05 (*)
Article Snippet: Transduction and selection of stable clones Myosin-X shRNA (m) Lentiviral Particles (Cat # sc-43,242-V), Control shRNA Lentiviral Particles-A (Cat # sc-108,080), Polybrene (Cat # sc-134,220) and puromycin dihydrochloride (Cat # sc-108,071) were purchased from Santa Cruz Biotechnology, Inc., and were used according to the manufacturer’s instructions.
Techniques: Western Blot, Expressing, Transfection, Plasmid Preparation, Infection, Activity Assay