*4.12. Generation of Conditioned Medium (CM) from Human M*Φ

Human monocytes were isolated from commercially available, anonymized buffy coats (DRK-Blutspendedienst Baden-Württemberg-Hessen, Frankfurt, Germany) using Ficoll-Hypaque gradients (PAA Laboratories, Cölbe, Germany) as previously described [41]. Briefly, monocytes were differentiated into primary human MΦ with RPMI-1640 containing 5% AB-positive human serum (DRK-Blutspendedienst Baden-Württemberg-Hessen, Frankfurt, Germany). Prior to stimulation, cells were serum-starved for 24 h and stimulated with 20 ng/mL IL-10 (Peprotech, Hamburg, Germany) for 24 h to generate conditioned-media of polarized MΦ [41]. Conditioned-media from iron-releasing MΦ were collected and used for following proliferation and migration assays. Supernatant of unstimulated MΦ served as control.

#### *4.13. Proliferation and Migration Assays*

Proliferation and migration assays were performed using the xCELLigence RTCA DP instrument (OLS, Bremen, Germany) as previously described [53]. Proliferation was recorded continuously for 3 days and migration for 24 h. Data were acquired as a measure for time-dependent impedance changes. RTCA Software 1.2 (OLS) was used for acquisition and analysis.
