Journal: International Journal of Molecular Sciences

Article Title: The Autophagy Nucleation Factor ATG9 Forms Nanoclusters with the HIV-1 Receptor DC-SIGN and Regulates Early Antiviral Autophagy in Human Dendritic Cells

doi: 10.3390/ijms24109008

Figure Lengend Snippet: DC-SIGN engagement induces autophagy and associates with LC3. ( A ) HEK293T cells were co-transfected with plasmids expressing DC-SIGN WT and GST-expressing constructs as indicated. Cells were engaged or not with DC-SIGN mAbs (5 μg/mL) for 30 min before pull-down. Lysates (input control) and GST pull-down were immunoblotted with anti-DC-SIGN ( upper panels), anti-GST ( middle panels), and anti-GAPDH for input control. This experiment is representative of 4. ( B ) Lysates from MoDC treated for indicated times with DC-SIGN specific mAbs were immunoblotted with anti-LC3, and loading was controlled with anti-GAPDH ( upper blots). Negative control experiments were performed using lysates of cells incubated with non-specific mouse serum (Isotype Ab) and immunoblotted as before ( lower blots). Pre-treatments of MoDC with chloroquine (50 μM) were performed to validate functional autophagy flux as evidenced by LC3-II increase. Densitometry values and LC3-II/GAPDH ratio obtained from lysates of primary MoDC treated with DC-SIGN mAbs (n = 6) or Isotype mouse Abs (n = 3) were obtained for each time point of each condition and graphically reported on the right. The same experiment as above was performed but with lysates from MoDC treated with ( C ) ManLAM (2 μg/mL) (n = 4) or ( D ) challenged with HIV-1-R5 (MOI of 2) (n = 3) for indicated times. LC3-II/GAPDH ratio from densitometry analyses was graphically represented as before. Statistical significance: * = p < 0.05; ** = p < 0.01.

Article Snippet: Chemiluminescence was acquired (Chemidoc, Bio-Rad Laboratories, Hercules, CA, USA), followed by densitometry analysis (Image Lab TM software; Bio-Rad Laboratories, Hercules, CA, USA).

Techniques: Transfection, Expressing, Construct, Control, Negative Control, Incubation, Functional Assay

Journal: International Journal of Molecular Sciences

Article Title: The Autophagy Nucleation Factor ATG9 Forms Nanoclusters with the HIV-1 Receptor DC-SIGN and Regulates Early Antiviral Autophagy in Human Dendritic Cells

doi: 10.3390/ijms24109008

Figure Lengend Snippet: ATG9 is required for DC-SIGN-mediated autophagy flux activation. Primary MoDC were treated with irrelevant siRNA (siCtrl) or siRNA against ATG9 (siATG9), and ATG9 expression was controlled by RT-qPCR ( left graph). Lysates from MoDC transfected as above and pre-treated with bafilomycin A1 (50 nM) for 1 h before stimulation with ManLAM (2 μg/mL) for 2 h were immunoblotted with anti-LC3 ( upper blot). The loading control was performed with anti-actin ( lower blot). LC3-II/actin ratio from densitometry analyses obtained from 3 independent experiments (n = 3) was normalized to untreated controls of each siRNA condition and graphically represented ( right graph). Statistical significance: * = p < 0.05.

Article Snippet: Chemiluminescence was acquired (Chemidoc, Bio-Rad Laboratories, Hercules, CA, USA), followed by densitometry analysis (Image Lab TM software; Bio-Rad Laboratories, Hercules, CA, USA).

Techniques: Activation Assay, Expressing, Quantitative RT-PCR, Transfection, Control

Journal: Veterinary radiology & ultrasound : the official journal of the American College of Veterinary Radiology and the International Veterinary Radiology Association

Article Title: Quantification of Nasal Airflow Resistance In English Bulldogs Using Computed Tomography and Computational Fluid Dynamics

doi: 10.1111/vru.12531

Figure Lengend Snippet: The process for generation of a 3D model. A: Computed tomographic images of the nasal passages were acquired in a transverse image plane using multi-detector computed tomography. A representative image of the caudal nasal passage at the level of the choanae is shown. B: Threshold segmentation in the ScanIP program isolated the airways (blue) using a threshold of −1024 to −450 Hounsfield units (HU). C: A fill threshold algorithm in ScanIP was used to generate a solid model of the nasal passages which was then sub-divided into 6-sided tetrahedral elements and exported to the COMOSL Multiphysics software package. D: 3D model of the nasal passages within the COMSOL Multiphysics package with E: Zoomed in image of the 3D model and tetrahedral mesh elements used in the computational fluid dynamic analysis. Note that each tetrahedral element has 6 sides or edges and the minimum edge length controls the mesh density where a smaller minimum edge length leads to a more dense mesh and more elements.

Article Snippet: First, the raw MDCT bone algorithm transverse dataset was imported into a three-dimensional (3D) image processing and model generation software program (ScanIP, Synopsys, Inc. Simpleware, Version 7.0, Chantilly, VA).

Techniques: Computed Tomography, Isolation, Software

Journal: Veterinary radiology & ultrasound : the official journal of the American College of Veterinary Radiology and the International Veterinary Radiology Association

Article Title: Quantification of Nasal Airflow Resistance In English Bulldogs Using Computed Tomography and Computational Fluid Dynamics

doi: 10.1111/vru.12531

Figure Lengend Snippet: Asymmetry of pressure between the right and left nasal passages was identified in two dogs. A: Rostrocaudal orientation; smoothed 3D model of the airway showing the right nasal cavity having a higher pressure compared to the left nasal passage. The yellow color depicts a higher pressure relative to the blue color-coded left nasal cavity. B: Ventrodorsal orientation; smoothed 3D model airway shows the differing pressures within the rostral nasal passages. The rostral nares are at the top of the image. C: Transverse CT image of a nasal cavity with asymmetry between the nasal cavities. Dog is in ventral (V) recumbency, the right side (R) of the dog is to the left of the image. Bone algorithm. Window width: 2500 Window level: 250.

Article Snippet: First, the raw MDCT bone algorithm transverse dataset was imported into a three-dimensional (3D) image processing and model generation software program (ScanIP, Synopsys, Inc. Simpleware, Version 7.0, Chantilly, VA).

Techniques: