Anti P75ntr, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Rabies Virus Hijacks and Accelerates the p75NTR Retrograde Axonal Transport Machinery"
Article Title: Rabies Virus Hijacks and Accelerates the p75NTR Retrograde Axonal Transport Machinery
Journal: PLoS Pathogens
Figure Legend Snippet: Suggested model. In order to arrive at the cell body and subsequently the CNS, rabies virus hijacks a fast route using the p75NTR endosomal pathway. In a p75NTR dependent path, RABV manipulates the axonal transport machinery to migrate faster to the cell body. An alternative, slower path, may involve alternative RABV receptors.
Figure Legend Snippet: RABV is retrogradely transported with neurotrophin receptors. ( A–D ), Retrograde transport of EGFP-RABV, added to the distal axon compartment of DRG explant previously treated with fluorescent antibodies against p75NTR and TrkA. Arrowheads: RABV puncta positive for p75NTR, arrows: RABV puncta positive for p75NTR and TrkA. Scale bar = 10 µm. ( E,F ) Co-localisation of RABV with p75NTR and TrkA calculated from two and one experiments, respectively. ( G ) Trajectories of RABV trafficked with neurotrophin receptors (NTFR, Blue) or without (Red), illustrating a more processive displacement over time of RABV with NTFR. ( H ) Merged kymographs of RABV (green) p75NTR (red) and TrkA (cyan), drawn for multi-channel time lapse. Vertical scale bar = 5 µm, horizontal scale bar = 40 seconds.
Figure Legend Snippet: RABV binds and internalizes with p75NTR in DRG neuron tips. Co-localization of EGFP-RABV with p75NTR is shown by live TIRF imaging and sub-pixel localization algorithms. ( A ) RABV-p75 particles shift from the periphery to the center of the growth cone, where they are internalized into the cell. Lower panels zoom in on dashed square, showing co-localized puncta (left) shifting towards the center of the growth cone (middle) until finally internalized (right). ( B ) Presentation of six separate events of RABV and p75NTR binding and internalization on the surface of the growth cone shown in (A). Colored trajectories denote displacement from point of detection to point of disappearance. ( C ) RABV and p75NTR are internalized together, illustrated by corresponding plots of puncta intensity over time (normalized to background), calculated for co-localized particles shown in lower panels of (A). Scale bars = 5 µm. ( D ) Zoom-in on colocalized RABV and p75 spot, taken from panel (A), scale bar = 1 µm. ( E ) Overlay of 1D-Gaussian fits of p75 and RABV intensity profiles at the x-axis of the image in panel (D). ( F ) Representative overlay of radial symmetry fits of the x-y intensity profiles of p75 and RABV spots. σ is the standard deviation of each fitting function; distance between the two spot centers is 51.3 nm. ( G ) Knockdown of p75NTR decreases rabies virus infection for shorts time incubation. DRGs embryonic cells infected with lentiviral vectors (LV) containing 4 different EGFP-tagged shRNA's against p75NTR or LV-EGFP, were transfected with RABV for 30 or 120 minutes. Low levels of infected neurons were found in shRNA-p75-EGFP cells Average RABV infection rates were normalized to LV-EGFP controls (n = 4 experiments, error bars = SEM, *p
Techniques Used: Imaging, Binding Assay, Standard Deviation, Infection, Incubation, shRNA, Transfection
Figure Legend Snippet: RABV travels faster and is more directed when transported with p75NTR. ( A–C ) Multi-channel live imaging of EGFP-RABV 2 hours after addition to distal axon compartment of DRG explant previously treated with a fluorescent antibody against p75NTR. Arrowheads: p75NTR-positive RABV puncta, scale bar = 10 µm. ( D,E ) Kymographs of and P75NTR extracted from time lapse depicted in (A–C). ( F ) RABV-only tracks (green) are less directed than RABV-p75NTR tracks (yellow), as shown when overlaying corresponding kymographs. Vertical scale bar = 5 µm, horizontal scale bar = 40 seconds. ( G–O ) Characterization of directed RABV puncta, transported with and without p75NTR, n = 184 and n = 122, respectively. (G) RABV presents higher speeds when transported with p75NTR, due to less frequent (H) and shorter pauses (I). Overall RABV-p75NTR spent less time paused on average (J), Diameter and intensity measurements revealed that RABV puncta positive for p75NTR were larger ( K ) and had higher intensity levels ( L ) than p75NTR-negative puncta. ( M–O ) p75NTR positive puncta (blue) are faster, more directed and present higher displacements over time, compared to p75NTR negative puncta (red), illustrated by distribution of instantaneous velocities in (M) (RABV+p75: n = 8051 events; RABV-p75: n = 7423 events) displacement plotted over time (N) and mean square displacement (O). Data is pulled from two separate experiments, error bars represent SEM. *p
Techniques Used: Imaging