Journal: Nature Communications

Article Title: Inflammation causes insulin resistance in mice via interferon regulatory factor 3 (IRF3)-mediated reduction in FAHFA levels

doi: 10.1038/s41467-024-48220-5

Figure Lengend Snippet: a Western blot of pAKT (S473) in WT SVF-derived adipocytes pre-treated with indicated conditioned medium plus insulin (100 nM) for 30 min. b Scheme showing the experimental paradigm used to identify IRF3 target genes by RNA-sequencing. Created with Biorender.com. c Volcano plot of RNA-seq dataset from ( b ). Genes that meet both significance and abundance are shown in black. Irf3 and Aig1 are marked in red. mRNA levels of Aig1 ( n = 3) ( d ) and western blot ( e ) of AIG1 in WT and FI3KO, FI3OE SVF-derived adipocytes. mRNA levels of Aig1 in iWAT, eWAT, and BAT from WT, FI3KO ( f ) and FI3OE mice ( g ) ( n = 5). h Western blot of AIG1 in iWAT, eWAT, and BAT of WT and FI3KO mice ( n = 3). i Western blot of AIG1 in iWAT, eWAT, and BAT of WT and FI3OE mice ( n = 3). Statistical significance was assessed by two-tailed Student’s t test. Data in all panels are expressed as mean ± SEM.

Article Snippet: Adeno-associated virus serotype 8 expressing mouse Aig1 and AAV8-GFP control were purchased from Applied Biological Materials (Canada).

Techniques: Western Blot, Derivative Assay, RNA Sequencing Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Inflammation causes insulin resistance in mice via interferon regulatory factor 3 (IRF3)-mediated reduction in FAHFA levels

doi: 10.1038/s41467-024-48220-5

Figure Lengend Snippet: a Proposed mechanism of covalent AIG1 inhibition by ABD-110. b Schematic illustrating the strategy for the AIG1 inhibitor treatment of WT and FI3OE mice on HFD at thermoneutrality. Figure created with Biorender.com. Insulin tolerance test ( c ), glucose tolerance test ( d ), serum insulin levels ( e ), in vivo 2-deoxyglucose uptake in BAT, iWAT, eWAT, quadriceps muscle, and heart ( f ) of mice described in ( b ) ( n = 7–8). g Quantification of FAHFA levels in WT and FI3OE SVF-derived adipocytes treated with or without ABD-110 (1 μM, 4 h) ( n = 12). Statistical significance was assessed by two-way ANOVA. Data in all panels are expressed as mean ± SEM. * P < 0.05 vs WT, # P < 0.05 vs vehicle.

Article Snippet: Adeno-associated virus serotype 8 expressing mouse Aig1 and AAV8-GFP control were purchased from Applied Biological Materials (Canada).

Techniques: Inhibition, In Vivo, Derivative Assay

Journal:

Article Title: Leukocyte-Mediated Endothelial Cell Injury and Death in the Diabetic Retina

doi:

Figure Lengend Snippet: CD18 blockade prevents leukocyte adhesion and endothelial cell injury and death in diabetic retinal venules and capillaries. A: After treatment with anti-CD-18 F(ab′)2, diabetic retinal leukocyte adhesion was markedly decreased in the retinal venules and capillaries. B: Analysis with PI revealed a concomitant decrease in retinal endothelial cell injury and death; however, no change in arteriolar endothelial cell injury and death was observed.

Article Snippet: In separate experiments, diabetic rats received either 1 mg/kg of a mouse anti-rat CD18 F(ab′) 2 (WT.3) (Associates of Cape Cod Inc., Falmouth, MA) or a purified mouse anti-human non-immune F(ab′) 2 (Jackson ImmunoResearch Laboratories Inc., West Grove, PA).

Techniques:

Journal:

Article Title: Leukocyte-Mediated Endothelial Cell Injury and Death in the Diabetic Retina

doi:

Figure Lengend Snippet: Anti-CD18 surface staining of a static leukocyte within a vessel. Green fluorescence from the FITC-linked Concanavalin A (A) and red fluorescence from the anti-CD18 antibody (B) identify the leukocytes as being CD18-positive when the images are superimposed (C).

Article Snippet: In separate experiments, diabetic rats received either 1 mg/kg of a mouse anti-rat CD18 F(ab′) 2 (WT.3) (Associates of Cape Cod Inc., Falmouth, MA) or a purified mouse anti-human non-immune F(ab′) 2 (Jackson ImmunoResearch Laboratories Inc., West Grove, PA).

Techniques: Staining, Fluorescence

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Development and characterization of xenograft model systems for adenoid cystic carcinoma

doi: 10.1038/labinvest.2011.105

Figure Lengend Snippet: Hematoxylin and eosin (H&E) stained histologic images of human (H) ACC tumors and their xenografts (X). The passage number (P) of the xenograft tumor being depicted is noted in the image. Similarly, representative images of immunohistochemistry (IHC) for versican and p63 are shown for the paired samples. In all instances, the xenograft tumors recapitulated the histologic and protein expression patterns of the human tumors from which they were derived. The immunohistochemistry was performed on tissue from the same passage number xenograft as depicted in the H&E images. (original magnification x400)

Article Snippet: A mouse monoclonal antibody to human large proteoglycan (versican) (clone# 2-B-1;Catalog# 270428-1; Associates of Cape Cod, Inc.) was used at 1:4000 dilution after microwave antigen retrieval.

Techniques: Staining, Immunohistochemistry, Expressing, Derivative Assay

Journal: Fertility and sterility

Article Title: Trophoblast retrieval and isolation from the cervix (TRIC) for noninvasive prenatal screening at 5 to 20 weeks of gestation

doi: 10.1016/j.fertnstert.2014.04.008

Figure Lengend Snippet: Expression of β-hCG in cells isolated using anti-HLA-G coupled to magnetic nanoparticles. Bound (A–C and G–I) and the excluded nonbound (D–F) cells were labeled with anti-β-hCG (A–F) or nonimmune IgG (G–I) and examined by immunofluorescence microscopy. Images of secondary antibody conjugated to FITC (green) appear in the left panels and matching fields of nuclei counterstained with DAPI (blue) appear in the center panels. Overlays of the FITC and DAPI stain appear in the right panels. Size bar is 50 μm.

Article Snippet: Immunomagnetic Isolation of Trophoblast Cells Mouse anti-HLA-G antibody (5 mg) was incubated overnight at 4°C in 100 mL sterile PBS containing 20 mL of 250 nm magnetic nanoparticles conjugated to goat anti-mouse IgG (Clemente Associates).

Techniques: Expressing, Isolation, Labeling, Immunofluorescence, Microscopy, Staining