Prolong gold mounting medium containing dapi

The Institutional Animal Care and Use Committee of University of Pittsburgh approved all experiments. The samples came from animals enrolled in completed studies. While in-study, animals were housed and maintained in accordance with standards established in the Animal Welfare Act and the Guide for the Care and Use of Laboratory Animals. The University of Pittsburgh is an AAALAS certified program. Non-human primate studies: Formalin-fixed paraffin-embedded (FFPE) tissue sections were cut from samples obtained from macaques as previously described [4 (link),7 (link)]. Antigen retrieval and staining were performed as previously described [5 (link),22 (link)]. Tissue sections were stained for vimentin (chicken polyclonal, 1:100 dilution; Novus Biologicals, Centennial, CO), α-smooth muscle actin (αSMA) (clone 1A4, 1:100 dilution; ThermoFisher Scientific, Waltham, MA), CD11c (clone 5D11, 1:30 dilution; Leica Microsystems, Buffalo Grove, IL), and CD31 (clone LCI-9, 1:50, Abcam, Cambridge, MA). Coverslips were applied using Prolong Gold mounting medium containing DAPI (ThermoFisher Scientific) and slides were imaged using a Nikon e1000 immunofluorescent microscope (Melville, NY) with a scanning stage. Images were saved as TIFF-format images, and processed with the Fiji build of ImageJ and the Template Matching plugin to align samples [25 (link),26 (link)].