Extended Data Fig. 6: Mitochondrial iron overload in macrophages during HFD promotes adipocyte iron overload and inflammation.
(a-q) Six-week-old male control and Mac-FtmtTG mice were fed HFD dox for 6 weeks. (a) Total iron content measured by ICP-MS in mitochondrial (n = 8) and cytosolic (control n = 4, TG n = 5) compartments in peritoneal macrophages. (b-d) Labile iron pool (LIP) in (b) ATMs (control n = 4, TG n = 5), (c) adipocytes (control n = 9, TG n = 12) and (d) SVF (control n = 7, TG n = 5) from eWAT. (e-f) mRNA levels of iron metabolism genes relative to control in (e) adipocytes (n = 5) and (f) ATMs (n = 6) from eWAT. (g) Ferritin (control n = 7, TG n = 12) and (h) Ferroportin (n = 3) protein expression relative to β-actin in adipocytes from eWAT. (i) Representative Western blots chosen from 3 independent experiments. (j) Ferritin (control n = 7, TG n = 9) and (k) Ferroportin protein expression relative to β-actin in ATMs from eWAT (n = 3). (l) Representative Western blots chosen from 3 independent experiments. (m) mRNA levels of mitochondrial iron metabolism genes relative to control in ATMs from eWAT (n = 6). (i) MitoNEET protein expression relative to β-actin in adipocytes from eWAT (control n = 4, TG n = 10). (o) LIP in mitochondrial and cytosolic compartments of eWAT (n = 4). (p-q) mRNA levels of inflammatory marker genes relative to control in (p) ATMs and (q) adipocytes from eWAT (n = 5). Significance in (a-h, j-k, m-q) between control and Mac-FtmtTG was calculated using a two-tailed Student’s t-test. Error bars represent mean ± S.E.M. * (P < 0.05), ** (p < 0.01), *** (p < 0.0001), **** (p < 0.00001).
