Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Leukocyte Chemotactic Receptor Fpr1 Protects Against Aging-related Posterior Subcapsular Cataract Formation

doi: 10.1096/fj.202002135R

Figure Lengend Snippet: (A) Enucleated eyes from Fpr1−/− and Fpr1+/+ mice at 7-months of age. Images are representative of at least 10 mice studied at this age. (B) Eye weight. Antero-posterior MRI (C) and ultrasound (D) images of the eye in 20-month old living mice. In (C), diameters are indicated in mm for the lens (yellow) and whole eye (red). In (D), severe cataracts and lens rupture are marked with the red and white arrows, respectively, the measurements of the eye diameters are in blue, and the labels for each compartment of the eye are in green. Images in C and D are representative of 3 Fpr1−/− mice and 3 Fpr1+/+ mice each that were scanned and analyzed. *p<0.05; **p<0.01; ***p<0.001, ns (not significant).

Article Snippet: The primers and probe used to detect human FPR1 were predesigned by the manufacturer (Applied Biosystems, Carlsbad, CA).

Techniques:

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Leukocyte Chemotactic Receptor Fpr1 Protects Against Aging-related Posterior Subcapsular Cataract Formation

doi: 10.1096/fj.202002135R

Figure Lengend Snippet: H&E stained antero-posterior sections of methacrylate-embedded eyes are shown for Fpr1+/+ and Fpr1−/− mice at the indicated ages. 200x magnification of the equatorial region (A) and posterior region (B) is shown to the right of each full eye image. (A) 5-month-old mice. Yellow arrows in the Fpr1−/− eye point to pyknotic nuclei in the mid equator region. (B) 7-month-old mice. Note the posterior plaque at the posterior pole and abnormal posterior migration of LECs in the Fpr1−/− lens (box). The red arrow in the magnified region points to one of the nuclei from an LEC in the posterior region. (C) Representative 20-month-old Fpr1+/+ mouse (left) and 7- and 20-month-old Fpr1−/− mice. The red arrow in the 7-month old lens points to the rupture at the posterior pole of the lens in an Fpr1−/− mouse. Data are representative of at least 5 mice in each group studied at 5, 7 and 20 months of age, respectively.

Article Snippet: The primers and probe used to detect human FPR1 were predesigned by the manufacturer (Applied Biosystems, Carlsbad, CA).

Techniques: Staining, Migration

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Leukocyte Chemotactic Receptor Fpr1 Protects Against Aging-related Posterior Subcapsular Cataract Formation

doi: 10.1096/fj.202002135R

Figure Lengend Snippet: (A and B) Human. Expression of FPR1 protein (A) and RNA (B) was analyzed for the indicated cell types by flow cytometry and real-time PCR, respectively. FACS plots show representative histograms for staining with the anti-FPR1 antibody (black) and isotype control antibody (grey). (C) Mouse. Fpr1 mRNA was quantitated by RT-PCR in the indicated tissue sources from 3-month-old wildtype C57Bl/6 mice. Data are representative of 2 independent experiments each for human and mouse lens and FHL124 cells.

Article Snippet: The primers and probe used to detect human FPR1 were predesigned by the manufacturer (Applied Biosystems, Carlsbad, CA).

Techniques: Expressing, Flow Cytometry, Real-time Polymerase Chain Reaction, Staining, Reverse Transcription Polymerase Chain Reaction

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Leukocyte Chemotactic Receptor Fpr1 Protects Against Aging-related Posterior Subcapsular Cataract Formation

doi: 10.1096/fj.202002135R

Figure Lengend Snippet: (A) Validation of goat anti-Fpr1 antiserum specificity by immunocytochemistry. The cell types analyzed are indicated at the upper left of each panel. HEK, parental or Fpr1-transfected human embryonic kidney 293 cells. Neutrophils are from mice with the indicated genotypes. In panel iii, the antiserum was pre-absorbed with the immunogenic peptide used to raise it. Isotype control antibody staining was negative (not shown). (B) Immunolocalization of Fpr1 in mouse lens. Whole eyes were transverse-sectioned through the optic nerve and pupil. Primary antibodies and mouse genotypes are indicated at the top right of each panel. Control IgG antisera were goat anti-FOXE3 (panel iii) and rabbit anti-PROX1 (panel vi). Anti-FoxE3 stained the cornea but not the lens (not shown), whereas we did not observe anti-PROX1 staining of any ocular structure under the conditions used. Staining was revealed with an appropriate secondary antibody conjugated to Alexa Fluor 594. Orange arrows indicate the anterior lens epithelial monolayer. Nuclei are counterstained blue with DAPI in each image in both A and B. Results are representative of at least 3 independent experiments.

Article Snippet: The primers and probe used to detect human FPR1 were predesigned by the manufacturer (Applied Biosystems, Carlsbad, CA).

Techniques: Immunocytochemistry, Transfection, Staining