Cathepsin b inhibitor 2 calbiochem

Cell damage and inflammatory markers were assessed within LLF of the mice. Cell damage was determined by release of lactate dehydrogenase (LDH; CytoTox 96 cytotoxicity assay, Promega; Madison, WI, USA) and protein concentration (Pierce BCA protein assay, Thermo Fisher Scientific; Waltham, MA, USA) according to manufacturer’s instructions. Cytokines were assessed by using a customized MesoScale Discovery U-PLEX assay platform (IFNγ, IL-1β, TNFα, IL-33, IL-6, IL-10, IL-13; Meso Scale Diagnostics LLC; Rockville, MD, USA) according to manufacturer’s instructions. Phagolysosomal membrane permeability (LMP) analysis was determined by total cathepsin (CTS; Z-LR-AMC fluorogenic peptide substrate, R&D Systems; Minneapolis, MN, USA) and cathepsin B (cathepsin B inhibitor II - Calbiochem, Millipore Sigma; Burlington, MA, USA) release within the LLF and IL-1β release (mouse IL-1β DuoSet ELISA, R&D Systems) within the supernatants of isolated ex vivo AM.

Cell damage and inflammatory markers were assessed within LLF of the mice. Cell damage was determined by release of lactate dehydrogenase (LDH; CytoTox 96 cytotoxicity assay, Promega; Madison, WI, USA) and protein concentration (Pierce BCA protein assay, Thermo Fisher Scientific; Waltham, MA, USA) according to manufacturer’s instructions. Cytokines were quantitated by using a customized MesoScale Discovery U-PLEX assay platform (IFNγ, IL-1β, TNFα, IL-33, IL-6, IL-10, IL-13; Meso Scale Diagnostics LLC; Rockville, MD, USA) according to manufacturer’s instructions. Phagolysosomal membrane permeability (LMP) analysis was determined by total cathepsin (CTS; Z-LR-AMC fluorogenic peptide substrate, R&D Systems; Minneapolis, MN, USA) and cathepsin B (cathepsin B inhibitor II—Calbiochem, Millipore Sigma; Burlington, MA, USA) release within the LLF and IL-1β release (mouse IL-1β DuoSet ELISA, R&D Systems) within the supernatants of isolated ex vivo AM.