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. 2021 Oct 4;35(12):917–950. doi: 10.1089/ars.2020.8234

FIG. 9.

FIG. 9.

Oxidation of hemoglobin by macrophages. (A) Macrophages were grown on coverslips and were exposed to oxyHb (10 μM). The supernatant of macrophages was removed after 1, 2, 4, and 8 h. FerrylHb content of supernatants was analyzed with Western blot. (B) Macrophages were grown on coverslips and were exposed to oxyHb (10 μM) for 24 h. Cells were stained with an anti-ferrylHb antibody with Alexa Fluor 488 secondary antibody for ferrylHb (green). Images were taken with Leica TCS SP8 gated STED-CW nanoscopy. Images were deconvolved using Huygens Professional software. (C) Macrophages were grown on coverslips and were exposed to oxyHb (10 μM) or ferrylHb (10 μM) for 12 and 24 h. Cells were stained with Hoechst 33258 for DNA (blue), an anti-ferrylHb antibody with Alexa Fluor 488 secondary antibody for ferrylHb (green). Images were taken with Leica TCS SP8 gated STED-CW nanoscopy. Images were deconvolved using Huygens Professional software. FerrylHb intensity of macrophages was calculated by ImageJ software. Scale bars shown in the images represent 8, 10 μm, and (B) 25 μm. *p < 0.05; ***p < 0.001. Color images are available online.