Journal: PLoS Neglected Tropical Diseases

Article Title: Proteomic Analysis of Urine Exosomes Reveals Renal Tubule Response to Leptospiral Colonization in Experimentally Infected Rats

doi: 10.1371/journal.pntd.0003640

Figure Lengend Snippet: The graph represents relative contribution of proteins to the variance between the Leptospira -infected and uninfected control rat urine exosomes. High value of VIP score indicates great contribution of the proteins to the group separation. The green and red boxes on the right indicate whether the protein concentration is increased (green) or decreased (red) in the exosome of the infected rat urine vs. uninfected rat urine samples. For higher n value, a VIP score of 1.5 is considered to enable discrimination between 2 phenotypes. Even with the low n (= 3) per group that is employed in this study, the VIP score of the top 3 proteins is higher than 3, increasing the confidence. Alanyl (membrane) aminopeptidase, also called CD13 is the top protein with a VIP score of 5.72.

Article Snippet: Antibody against alanyl aminopeptidase was purchased from Proteintech Group, Inc., (Chicago, IL, USA).

Techniques: Infection, Control, Protein Concentration, Membrane

Journal: PLoS Neglected Tropical Diseases

Article Title: Proteomic Analysis of Urine Exosomes Reveals Renal Tubule Response to Leptospiral Colonization in Experimentally Infected Rats

doi: 10.1371/journal.pntd.0003640

Figure Lengend Snippet: Top discriminators between control, infected male & infected female rat urine exosome proteins.

Article Snippet: Antibody against alanyl aminopeptidase was purchased from Proteintech Group, Inc., (Chicago, IL, USA).

Techniques: Control, Infection, Membrane, Binding Assay

Journal: PLoS Neglected Tropical Diseases

Article Title: Proteomic Analysis of Urine Exosomes Reveals Renal Tubule Response to Leptospiral Colonization in Experimentally Infected Rats

doi: 10.1371/journal.pntd.0003640

Figure Lengend Snippet: Male infected versus Control.

Article Snippet: Antibody against alanyl aminopeptidase was purchased from Proteintech Group, Inc., (Chicago, IL, USA).

Techniques: Infection, Control, Membrane, Protease Inhibitor, Binding Assay

Journal: PLoS Neglected Tropical Diseases

Article Title: Proteomic Analysis of Urine Exosomes Reveals Renal Tubule Response to Leptospiral Colonization in Experimentally Infected Rats

doi: 10.1371/journal.pntd.0003640

Figure Lengend Snippet: Female infected versus Control.

Article Snippet: Antibody against alanyl aminopeptidase was purchased from Proteintech Group, Inc., (Chicago, IL, USA).

Techniques: Infection, Control, Membrane, Protease Inhibitor

Journal: PLoS Neglected Tropical Diseases

Article Title: Proteomic Analysis of Urine Exosomes Reveals Renal Tubule Response to Leptospiral Colonization in Experimentally Infected Rats

doi: 10.1371/journal.pntd.0003640

Figure Lengend Snippet: Proteins significantly dysregulated between exosomes from urines of control rats and infected rats.

Article Snippet: Antibody against alanyl aminopeptidase was purchased from Proteintech Group, Inc., (Chicago, IL, USA).

Techniques: Control, Infection, Membrane, Binding Assay, Protease Inhibitor

Journal: EMBO Molecular Medicine

Article Title: Photoreceptor glucose metabolism determines normal retinal vascular growth

doi: 10.15252/emmm.201707966

Figure Lengend Snippet: The mouse neonates received intraperitoneal injection of STZ and schematic graph shows mouse retinal vessel development after birth (left). GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer. Representative images of the deep vascular network in isolectin‐stained (red, top) retinal whole mounts of HAR and controls. Scale bar, 50 μm (top) and 1 mm (bottom). The total retinal area is outlined in yellow and deep vascular coverage is highlighted in red (bottom, grayscale). Quantification of deep vascular network of HAR and controls. n = 17–27 retinas/group. Data presented as mean ± SEM, unpaired t ‐test. Blood glucose ( n = 19–27 retinas/group), serum insulin (ELISA) ( n = 4–7 pooled samples/group) and triglyceride (TG) concentrations ( n = 7/group) of HAR and controls. Data presented as mean ± SEM, unpaired t ‐test. Data information: See also Fig .

Article Snippet: All serum samples were diluted 1:306 and APN levels assayed using a human adiponectin ELISA kit (E091M, Mediagnost, Reutlingen, Germany).

Techniques: Injection, Staining, Enzyme-linked Immunosorbent Assay

Journal: EMBO Molecular Medicine

Article Title: Photoreceptor glucose metabolism determines normal retinal vascular growth

doi: 10.15252/emmm.201707966

Figure Lengend Snippet: A Left: serum APN levels (ELISA) ( n = 8–9 retinas/group); right: retinal apn (qRT–PCR) ( n = 6 retinas/group) of HAR and controls. B Left: retinal cross‐sectional layers for laser capture microdissection (LCM: DAPI for nuclei, blue; isolectin for vessels, red); center: mRNA levels of AdipoR1 and AdipoR2 in retinal neuronal layers and vessels. n = 3 pooled retinas/group. Right: AdipoR1 mRNA in HAR and control retinas. n = 5 retinas/group. C Control and HAR eyes in WT and Apn −/− mice. Left: representative images of DAPI‐stained hyaloid vessels (blue). Right: quantification of preserved hyaloid vessels branching from the hyaloid artery (white arrow). Scale bar, 1 mm. n = 8–30 retinas/group. D Left: Fundus photograph focused to show persistent hyaloid vessels (white arrows) (green, fluorescein AK‐FLUOR) in WT and Apn −/− mice at P30 ( n = 37–48 retinas/group); right: pie graph of percentage of eyes examined with persisting hyaloid. E Left: In WT and Apn −/− representative images of deep retinal vasculature (lectin, red) in whole‐mounted retinas; right: quantification of deep vessels ( n = 14–22 retinas/group). F, G Left: Representative images of deep retinal vasculature (lectin, red) in whole‐mounted retinas of WT hyperglycemic (HAR) mice with recombinant mouse APN treatment (F) ( n = 6–10 retinas/group) or with AdipoRon treatment (G). Right: quantification of deep retinal vasculature ( n = 7–14 retinas/group). Data information: Scale bars, 50 μm (B, E–G) or 1 mm (C). Data presented as mean ± SEM; unpaired t ‐test (A, B, E–G) or ANOVA (C). See also Fig .

Article Snippet: All serum samples were diluted 1:306 and APN levels assayed using a human adiponectin ELISA kit (E091M, Mediagnost, Reutlingen, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Laser Capture Microdissection, Control, Staining, Recombinant

Journal: Gene Therapy

Article Title: A macrophage-specific synthetic promoter for therapeutic application of adiponectin

doi: 10.1038/gt.2014.3

Figure Lengend Snippet: Constructs of plasmid vectors for APN expression. ( a ) SP146-C1 was selected to develop a therapeutic vector. The original luciferase gene in the pGL3-basic plasmid was replaced by the APN gene. ( b ) Cells were transfected and mRNA expression of APN with CMV-APN and SP146-C1-APN (SP-APN) was detected 24 h later by RT-PCR in 293 T and RAW264.7 cells. CMV-APN was used as a positive control. Densities of bands were measured using Alpha easy FC software and normalized to that obtained with actin or glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( c ) Concentrations of APN in the media were measured 48 h later using a human APN ELISA Kit in 293 T and RAW264.7 cells. Data shown are means±s.d.'s. ( d ) The ratio of expressions of SP-APN and CMV-APN was calculated and expressed in graphs. Both mRNA level of APN and released APN showed high ratio in RAW264.7 cells. ** P <0.01 and *** P <0.001. APN, adiponectin; ELISA, enzyme-linked immunosorbent assay; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Cells were transfected with APN-encoding plasmid vector (CMV-APN) or APN-encoding SP plasmid vector (SP-APN) and the concentration of APN in media was determined using a human APN ELISA Kit (AbFrontier, Seoul, Korea) 48 h later according to the manufacturer's manual.

Techniques: Construct, Plasmid Preparation, Expressing, Luciferase, Transfection, Reverse Transcription Polymerase Chain Reaction, Positive Control, Software, Enzyme-linked Immunosorbent Assay