Figure 3.

Figure 3.

TMEM187 deficiency shifts iron metabolism, favoring erythroid differentiation in K562 cells. (A) Pathways involved in metal ion metabolism and hematopoiesis regulation were enriched by GO enrichment analysis before induction. (B-D) Absolute iron content, revealed by inductively coupled plasma-mass spectrometry (B), nonheme total iron content, revealed by ferrozine assays (C), and the relative cytosolic and mitochondrial labile iron pool (supplemental Methods) (D). t test was used for significance. ∗P < .05; ∗∗P < .01. (E-F) Protein levels of IRP2, TfR1, FTH, and FTL (E), quantified in (F). GAPDH was used as an internal control. (G-H) Levels of Fe-S cluster biosynthesis–related proteins (G), quantified in (H). GAPDH was used as an internal control. (I) The relative activities of FECH in WT and KO cells. (J) The heme content in WT and KO cells detected by heme enzyme-linked immunosorbent assay. One-way ANOVA was used for significance unless otherwise specified. n = 8/group for (C), n = 4/group for (J), others, n = 3/group. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. GO, Gene Ontology; ns, no significance.