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. Author manuscript; available in PMC: 2022 May 1.
Published in final edited form as: J Immunol Regen Med. 2021 Feb 2;12:100038. doi: 10.1016/j.regen.2021.100038

Figure 3: Macrophages ingest apoptotic ECs in wounds in vivio.

Figure 3:

pHrodo green labeled apoptotic mECs were injected into day 6 mouse skin wounds. The uptake of apoptotic mECs 24 hours after injection was examined. A) Images taken using a regular fluorescence microscope. B) Confocal microscopy images. The white arrow in Z stack images at the left panel indicates a macrophage with engulfed pHrodo green labeled apoptotic mEC. The 3D image at the right panel shows the pHrodo green labeled apoptotic mECs are embedded among red stained CD68 macrophages suggesting macrophages ingested apoptotic mECs. C) Cells were isolated from the wounds on day 7 and subjected to flow cytometry to identify F4/80 macrophages that had ingested apoptotic mECs. Cells were collected from 8 pooled wounds from 4 mice (two wounds from each mouse) in each group. The number shown in the right scatter plot is the percentage of F4/80 macrophages with engulfed pHrodo green labeled apoptotic mECs. Representative results from two experiments are shown. Gating strategies are shown in Appendix C