Distinct Lysosome Phenotypes Influence Inflammatory Function in Peritoneal and Bone Marrow-Derived Macrophages

Figure 1

Peritoneal macrophages have increased lysosome volume and activity compared to BMDMs. (a, b) Thioglycollate-elicited peritoneal macrophages (pMACs) or bone marrow-derived macrophages (BMDMs) were stained with lysotracker red and analyzed by flow cytometry. A representative histogram (a) and FL2 mean fluorescence intensity (MFI) quantification (b) are shown. (c, d) Total LAMP1 was determined by staining fixed and permeabilized pMACs or BMDMs with a LAMP1 (CD107a) PE antibody followed by flow cytometry assessment. A representative histogram (c) and MFI quantification (d) are shown. (e, f) Cathepsin B activity was assessed using a fluorogenic cathepsin B substrate that requires cleavage to produce fluorescent signal. After incubation with substrate, pMACs and BMDMs were analyzed by flow cytometry. A representative histogram (e) and MFI quantification (f) are shown. Bar graphs report the mean ± standard error (SE) for a minimum of 3 experiments, each performed in triplicate. for pMAC versus BMDM.

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