Journal: Genes

Article Title: Human Vascular Endothelial Growth Factor A 165 Expression Induces the Mouse Model of Neovascular Age-Related Macular Degeneration

doi: 10.3390/genes9090438

Figure Lengend Snippet: Morphologic changes and transgene expression in the Cre and LacZ injected eyes. ( a ) β-gal expression (arrowhead) after LacZ injection was seen two weeks after gene transfer but not at later time points. ( b ) VEGF-A expression (violet) in ganglion cell layer (black arrowhead), photoreceptors (arrowhead), and neovascular membrane (arrow) in the eye of Cre -injected mouse. ( c ) Glial fibrillary acidic protein (GFAP) immunoreactivity was observed in the nerve fiber layer (arrowhead) and Müller cells (arrow) in the outer retina post- Cre injection. ( d ) In the Cre group, F4/80 positive macrophages were seen in the retina and subretinal layers. ( e , f ) Retinal autofluorescence (yellow) with DAPI nuclear counterstain (blue). In Cre -injected retina (e), drusen-like lipofuscin deposits (arrowhead) and the loss of photoreceptors were seen. Intact photoreceptor layer (f, arrowhead) was observed in LacZ -injected eyes. Scale bar is 100 µm. GCL: Ganglion cell layer.

Article Snippet: For immunostainings, the following antibodies were used: CD34 (MEC14.7, Hycult Biotech, Uden, The Netherlands), glial fibrillary acidic protein (GFAP) (Z0334, Dako, Santa Clara, CA, USA), F4/80 (MCA497R, Bio-Rad, Hercules, CA, USA), β-gal (AB1211, EMD Millipore, Billerica, MA, USA), and VEGF (ab52917, Abcam).

Techniques: Expressing, Injection

Journal: Investigative Ophthalmology & Visual Science

Article Title: Corneal Neurotoxicity Due to Topical Benzalkonium Chloride

doi: 10.1167/iovs.11-8775

Figure Lengend Snippet: Corneal inflammation, NFD, and aqueous tear production after 1 week of treatment with BSS or BAK. (A) Hematoxylin–eosin staining of corneas treated with BSS are compact with no inflammation. (B) Corneas treated with 0.1% BAK have corneal edema and inflammatory cells (arrowhead points to neutrophils). (C) Whole mount confocal immunostaining with CD11b and F4/80 antibody for inflammatory cells show normal nerves and the absence of inflammatory cells in BSS-treated corneas. (D and E). In 0.1% BAK-treated corneas, nerve fluorescence is lost and inflammation is present The YFP+ inflammatory cells in D and E (green), CD11b antibody (D, red), and F4/80 antibody (E, red) colocalize (D and E, arrowheads). (F) The YFP+ cells are present in the bone marrow as yellow fluorescent spheres. (G) Immunostaining with leukocyte common antigen (CD45) antibody (G, red) confirmed that YFP+ cells are of hematopoietic lineage. (H) Graph shows that corneal NFD was significantly less after 1 week of 0.01% BAK treatment and 0.1% BAK, but there was no change in NFD with BSS treatment. (I) Graph shows that YFP+ inflammatory cells were significantly increased after 1 week of 0.01% BAK treatment and 0.1% BAK. The inflammatory cells in normal and BSS-treated corneas were less that 10 cells/cornea, therefore the bars are not visible in the graph. (J) Graph shows that aqueous tear production was significantly less after 1 week of 0.01% and 0.1% BAK, but there was no change in tear production with BSS treatment. *P < 0.05, White scale bar for C, D, and E = 20 μm, F = 50 μm, and G = 10 μm.

Article Snippet: Primary antibodies used were Neurofilament H Non-Phosphorylated Monoclonal Antibody (SMI-32; catalog no. SMI-32R, Covance Inc., Princeton, NJ; antibody specificity for axonopathy validated by Bannerman and Hahn 20 and Irvine and Blakemore 21 ), rat anti-mouse F4/80 and CD11b (F4/80; catalog no. MCA 497R and CD11b; catalog no. MCA711 both from AbD Serotec, Raleigh, NC).

Techniques: Staining, Immunostaining, Fluorescence

Journal: Scientific Reports

Article Title: The pharmaceutical solvent N-methyl-2-pyrollidone (NMP) attenuates inflammation through Krüppel-like factor 2 activation to reduce atherogenesis

doi: 10.1038/s41598-020-68350-2

Figure Lengend Snippet: NMP treatment reduces macrophages infiltration and inflammation. ( a ) Representative immunostaining of macrophages, smooth muscle cells (SMC) and activated endothelium in aortic sinus lesions of NMP treated and untreated C57BL/6 J mice fed with HFD for 12 weeks. Lesions were stained for biomarkers of macrophages (CD68; yellow), SMCs (α-actinin; green) and stressed endothelium (Vcam-1; red); nuclei were stained with DAPI (blue). Merged images are also shown. Lm, aortic lumen, M, tunica media, and I, tunica intima. White dashed lines demarcate the elastic lamina. Bars, 200 μm. ( b ) Vcam-1 and merge staining as in ( a ) from different animals. ( c ) Quantitative analysis of relative Vcam-1 immunostaining relative to the aortic sections size from ( b ).

Article Snippet: Immunofluorescence samples were stained with rat anti-mouse CD68 F4/80 (AbD Serotec, MCA497R) and Cy3-conjugated mouse anti-smooth muscle actin (SMA) (Sigma-Aldrich, C6198).

Techniques: Immunostaining, Staining