Fig. 5: TIPE2-deficient macrophages drive ASC ferroptosis by propagating mitochondria fragmentation via TNTs. | Nature Communications

Fig. 5: TIPE2-deficient macrophages drive ASC ferroptosis by propagating mitochondria fragmentation via TNTs.

From: Obesity-associated macrophages dictate adipose stem cell ferroptosis and visceral fat dysfunction by propagating mitochondrial fragmentation

Fig. 5

a, b Flow cytometry frequency (a) of mitochondrial ROS in ASCs with MFI quantification (b; n = 4 or 5) after 24 h coculture with Tipe2+/+ or Tipe2-/- macrophages. c, d Flow cytometry frequency (c) of lipid ROS in ASCs with MFI quantification (d; n = 4 or 5) after 24 h coculture with Tipe2+/+ or Tipe2-/- macrophages with or without 20 μM MitoTEMPO. e-g, Schematic diagram of coculture assay (e) for ASCs (wild-type) and MitoSOX-labeled macrophages (Tipe2+/+ or Tipe2-/-) in the presence of FA, and flow cytometry frequency (f) of macrophage-derived mitochondrial ROS in ASCs with MFI quantification (g; n = 3) after 24 h coculture. h–j Schematic diagram of coculture assay (h) for ASCs (wild-type) and MitoTracker-labeled macrophages (Tipe2+/+ or Tipe2-/-) in the presence of FA, and representative confocal imaging of macrophage-derived mitochondria in ASCs after 24 h coculture (i), or in the presence 350 nM cytochalasin B (CytoB) (j). Scale bars, 50 μm. k Representative confocal imaging of TNTs (F-actin, yellow) between ASCs and Tipe2-/- macrophages (F4/80, green) in the above coculture system and their co-localization with macrophage-derived mitochondria (red, MitoTracker). Nuclei (blue, DAPI). Scale bars, 50 μm. l–o Flow cytometry frequencies of lipid ROS with MFI quantification (l, m; n = 3 or 4) and cellular death (n, o; n = 4 or 5) in ASCs after 24 h coculture with Tipe2+/+ or Tipe2-/- macrophages in presence or absence of 50 μM Mdivi-1. Data are presented as mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001 determined by two-tailed student’s t-test (b,g) or one-way ANOVA (d,m,o). Data are representative of at least two independent experiments.

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