Fig. 2

Ferroptotic THP-1 cells promote the efferocytosis of LPMs in vitro. (A) Cell viability, (B) LDH relative content, and (C) the MDA concentration in THP-1 cells stimulated with different concentrations of Erastin for 24 h. (D) The intracellular ROS level stimulated with 80 µM Erastin for 24 h was measured by confocal microscope. (scale bar: 50 μm) (E) qRT-PCR analysis showed the mRNA level of Ptgs2 in THP-1 cells stimulated with 80 µM Erastin for 24 h (normalised to β-actin). (F) Representative pictures acquired by transmission electron microscopy. Red arrows indicated mitochondrial cristae disappearance and outer membrane rupture. (G) THP-1 cells treated with or without Erastin for 24 h, then stained by CFSE. MFI of CFSE in THP-1 cells was detected by flow cytometry. LPMs were incubated with nontreated or Erastin-treated THP-1 cells stained by CFSE for 4 h. (H, I) Efferocytosis of LPMs from each group was determined by (H) flow cytometry and (I) confocal microscope (scale bar: 50 μm). (J, K) LPMs pretreated with 5 µM CytoD for 1 h were incubated with nontreated or Erastin-treated THP-1 cells stained by CFSE for 4 h. Efferocytosis of LPMs from each group was determined by (J) flow cytometry and (K) confocal microscope (scale bar: 50 μm). (L) LPMs were incubated with nontreated or Erastin-treated THP-1 cells for 4 h. The mRNA levels of efferocytosis related genes in LPMs were measured by qRT-PCR (normalised to β-actin). Values are mean ± SD. *p < 0.05, **p < 0.01, ****p < 0.0001, ns denotes p > 0.05 (by unpaired Student’s t test)