Extended Data Fig. 3: Macrophages respond to mitoNEET overexpression by up-regulating iron storage.
(a-q) Male control and Mac-MitoNEETTG mice were fed chow dox for 2 weeks. (a) Labile iron pool (LIP) in mitochondrial and cytosolic compartments of peritoneal macrophages (control n = 7, TG n = 6). (b) mRNA levels of iron metabolism genes relative to control in peritoneal macrophages (control n = 10, TG n = 7). (c) Ferritin (control n = 9, TG n = 12), (d) Ferroportin (control n = 3, TG n = 5), (e) CD163 (control n = 3, TG n = 5), (f) CD91 (control n = 3, TG n = 5) and (g) FTMT (control n = 3, TG n = 5) protein expression relative to β-actin in peritoneal macrophages. (h) Representative Western blots chosen from 2 independent experiments (i) mRNA levels of inflammation markers relative to control in peritoneal macrophages (control n = 7, TG n = 10). (j-l) Flow cytometric measurement of total (control n = 6, TG n = 5), M1-like control n = 6, TG n = 5) and M2-like macrophages in eWAT (n = 6). (m) Body weight (control n = 3, TG n = 5). (n-o) Oral glucose tolerance test (control n = 3, TG n = 5). (n) Blood glucose levels. (o) Serum insulin levels. (p) Blood glucose levels during insulin tolerance test (control n = 3, TG n = 5). (q) Serum triglyceride levels during triglyceride clearance (n = 6). Significance in (a-g, i-m) between control and Mac-MitoNEETTG was calculated using a two-tailed Student’s t-test. Significance in (n-q) was calculated using a 2-way ANOVA with Sidak’s post-test for multiple comparisons. Error bars represent mean ± S.E.M. * (P < 0.05), ** (p < 0.01), *** (p < 0.0001), **** (p < 0.00001).
