benzyloxycarbonyloxy succinimide cbz osu  (Chem Impex International)

Journal: Disease Models & Mechanisms

Article Title: HTRA1-dependent proteolysis induces age-related retinal degeneration and exacerbates choroidal neovascularization

doi: 10.1242/dmm.052253

Figure Lengend Snippet: HTRA1 protein accumulates in lesions present in human donor eyes with age-related macular degeneration (AMD). Immunohistochemistry (IHC) showing HTRA1 distribution in retinas from unaffected controls, and patients with intermediate AMD (iAMD) and geographic atrophy (GA). Some retinas present macular neovascularization (MNV). (A-C) In control retinas, HTRA1 predominantly localizes to BM, the outer plexiform layer (excluding the Henle fiber layer) and horizontal cells, with variable, weaker staining of the photoreceptor layer. (D-G) Intermediate AMD retinas show HTRA1-positive basal laminar deposits, basal linear deposits and hard drusen between the RPE and BM, and accentuation of HTRA1 is seen at the RPE/photoreceptor interface, demonstrated in the magnification of D (G). (H-K) GA retinas show strong HTRA1 positivity in heavily pigmented cells associated with the degenerating RPE layer, which may represent displaced RPE cells or macrophages. (L) Parallel slide to K, showing CD31 IHC to confirm neovascularization. Filled arrowheads, BM; empty arrowheads, external limiting membrane descent; green arrowheads, neovascularization (vessels between BM and RPE). Each image is from a unique patient, except F and I, which are different eyes from the same patient, and K and L, which are from parallel sections from the same eye of a patient. Scale bars: 50 µm. RGC, retinal ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; RPE, retinal pigment epithelium; BM, Bruch's membrane.

Article Snippet: The sections were blocked with 3% bovine serum albumin, followed by 5 μg/ml mouse anti-human HTRA1 (Genentech, clone 4B1) or rabbit anti-human CD31 (Bethyl Labs, polyclonal) for 60 min. Signal was detected with Powervision polymer-AP anti-Ms (Leica Biosystems, PV6110) for 30 min and visualized with Fast Red (ScyTek, FR0001-1FU).

Techniques: Immunohistochemistry, Control, Staining, Membrane

Journal: Disease Models & Mechanisms

Article Title: HTRA1-dependent proteolysis induces age-related retinal degeneration and exacerbates choroidal neovascularization

doi: 10.1242/dmm.052253

Figure Lengend Snippet: Human HTRA1 overexpression in mice exacerbates laser-induced choroidal neovascular lesions. (A) Diagram of the laser-induced choroidal neovascularization (CNV) preclinical model. Four laser burns per eye were performed in No Tg mice and mice expressing catalytic dead HTRA1 (S328A, hemi) or catalytic active HTRA1 (WT, hemi). CNV lesions were allowed to grow for 1 week, then lesion surface area was quantified using vascular labelling (lectin perfusion and anti-CD31 immunofluorescence). Created in BioRender by Gu, X. (2025). https://biorender.com/muizvw7 . This figure was sublicensed under CC-BY 4.0 terms. (B) Representative immunofluorescence images showing lectin (red) and anti-CD31+anti-Emcn (green) labelling of four CNV lesions in an eye from a No Tg, a HTRA1 S328A or a HTRA1 WT-expressing mouse. Scale bars: 200 µm. (C) CNV lesion surface area quantification. N =8 to 9 mice per genotype (total of 46 to 62 lesions). One-way ANOVA; n.s., not significant ( P >0.05); **** P <0.0001.

Article Snippet: The sections were blocked with 3% bovine serum albumin, followed by 5 μg/ml mouse anti-human HTRA1 (Genentech, clone 4B1) or rabbit anti-human CD31 (Bethyl Labs, polyclonal) for 60 min. Signal was detected with Powervision polymer-AP anti-Ms (Leica Biosystems, PV6110) for 30 min and visualized with Fast Red (ScyTek, FR0001-1FU).

Techniques: Over Expression, Expressing, Immunofluorescence