VAMP3/Cellubrevin Antibody NB300-510 from Novus Biologicals, a Bio-Techne Brand

Published Figures
Supplier Images

Register or Sign In to View Figures.

Scientific reports
October 27 2021

Fig 1: Subcellular localization of endocytic BDNF in cultured astrocytes. (A) Representative fluorescence images of the colocalization of QD-BDNF with endogenous vesicular markers. ChgB: chromograninB. Scale bar = 2 µm. White arrowheads: representative colocalization of QD-BDNF with the corresponding markers. (B) Average colocalization ratios (# colocalized QD-BDNF/# total QD-BDNF). Dotted line: average colocalization ratio between QD-BDNF and MitoTracker (Mito.; negative control). **P < 0.01 (Vamp3 vs. others), ##P < 0.01 (Mito. vs. others). N = 39–45 cells (1,834–10,531 QD particles) for vesicular markers. (C) Representative fluorescence images of the colocalization of QD-BDNF, Vamp3-EGFP, and other vesicular markers. Scale bar = 2 µm. White arrowheads: representative triple colocalization among QD-BDNF, Vamp3-EGFP, and the corresponding vesicular markers. (D) Above: average colocalization ratio between Vamp3-EGFP and each vesicular marker. *P < 0.05, **P < 0.01. Below: average colocalization ratio of each vesicular marker with QD-BDNF with Vamp3-EGFP (Vamp3( +)) or without Vamp3-EGFP (Vamp3(-)). **P < 0.01. N = 9–10 cells (QD-BDNF particles: 124–481; Vamp3-EGFP puncta: 449–626). Colocalization ratios were determined by using ImageJ/FIJI software (Ver. 2.1.0/1.53c, NIH).
Register or Sign In to View Figures.

Molecular metabolism
November 01 2015

Fig 2: TUSC5 promotes intracellular associations during protein recycling. (A) Western blot of immunoprecipitation of endogenous cellugyrin (CG) from 3T3-L1 adipocytes after siRNA-mediated TUSC5 knockdown. (B) Western blot of immunoprecipitation of endogenous GLUT4 from 3T3-L1 adipocytes in control and TUSC5 knockdown conditions. (C) MS1 intensities from mass spectrometric analysis of co-precipitants of GLUT4 IPs as in 2D (n = 4). *p < 0.05, **p < 0.01 by Student's t-test. Bar graphs shown are mean ± s.e.m. (D) Representative confocal microscopy images of insulin-stimulated 3T3-L1 adipocytes (0, 15, 60 and 90 min stimulation after starvation) after siRNA-mediated TUSC5 knockdown. Quantification of (E) VAMP3 and (F) GLUT4 signal intensities at the plasma membrane from representative confocal microscopy images as shown in 4D. (G) Amount of surface exposed Glut4myc at the indicated time points in basal and insulin-stimulated L6 and L6Glut4myc cells after adenoviral mediated TUSC5 overexpression. *p < 0.05 T5 OE vs. scr ctrl by Student's t-test.
Register or Sign In to View Figures.

Scientific reports
October 27 2021

Fig 3: Vamp3 is necessary for ATP-induced endocytic BDNF secretion. (A) Representative QD-BDNF kymographs generated by using ImageJ/FIJI software (Ver. 2.1.0/1.53c, NIH), from astrocytes with GFP- (CTL), siSCR-, siVamp3-, or siVamp3 + human Vamp3 (siVamp3 + rescue). White arrowheads: disappearance of QD-BDNF particles. (B) Average intracellular QD-BDNF densities under each condition. N = 9–18 cells. The quantification of QD-BDNF particle numbers was determined by using ImageJ/FIJI software (Ver. 2.1.0/1.53c, NIH). (C) Average QD-BDNF fractions showing immobility (St) or anterograde (An)/retrograde (Re) transport. *P < 0.05, **P < 0.01, ****P < 0.0001. N = 10–16 cells. (D) Cumulative distributions of the QD-BDNF transport distances at baseline and after ATP stimulation (ATP) in each group. ****P < 0.0001. (E) Average velocities of QD-BDNF transport. N = 165 and 302 particles for the siSCR and siVamp3 groups, respectively. F. Average percentages of QD-BDNF secretion events after the indicated treatments. **P < 0.01, ***P < 0.001, ****P < 0.0001. N of tested cells (with QD particle number): naïve = 6 (102), siSCR = 12 (197), siVamp3 = 16 (304), siVamp3 + rescue = 13 (187).
Register or Sign In to View Figures.

Scientific reports
October 27 2021

Fig 4: ATP-induced secretion of endocytic BDNF from Vamp3-containing vesicles. (A) Representative fluorescence images of astrocytic processes containing QD-BDNF and Vamp3-EGFP. The kymograph was generated by using ImageJ/FIJI software (Ver. 2.1.0/1.53c, NIH). Black arrow heads: Vamp3-positive QD-BDNF particles. Empty arrowheads: Vamp3-negative QD-BDNF particles. Red bar: ATP treatment. White sharp arrowheads: disappearance of QD-BDNF particles. Black bar = 30 s. (B) Average percentages of ATP-induced QD-BDNF secretion events from vesicles with ( +) or without Vamp3 (−). **P < 0.01, ***P < 0.001. N = 15 cells (vehicle = 162; ATP = 333 QD particles). (C) Average fractions of secreted QD-BDNF particles with ( +) or without Vamp3 (−) among total secreted QD-BDNFs. ****P < 0.0001. N = 15 cells (107 secreted QD particles).