Fig. 5: Lysosomal lipid peroxidation was related to ROS but independent of the direct transport of LPO by SCP2.
From: SCP2 mediates the transport of lipid hydroperoxides to mitochondria in chondrocyte ferroptosis
A The merged images of BODIPY 665/676, CellLight-lyso-GFP, and Hoechst 33258 staining in rat chondrocytes treated with RSL3 (0.25 μM), 4OHT (1 μM), ScpI2 (5 μM), MitoQ (1 μM) or CQ (20 μM). Scale bar, 50 μm. B The relative colocalization coefficients of lysosomes and LPO with lyso-GFP and BODIPY staining (n = 6 per group). C The merged images of immunofluorescence SCP2, CellLight-lyso-GFP, and Hoechst 33258 staining in rat chondrocytes treated with RSL3 (0.25 μM), 4OHT (1 μM), ScpI2 (5 μM), MitoQ (1 μM) or CQ (20 μM). Scale bar, 50 μm. D The relative colocalization coefficients of lysosomes and SCP2 with lyso-GFP and immunofluorescence staining (n = 6 per group). E Lysosomal membrane permeabilization of rat chondrocytes treated with RSL3 (0.25 μM), 4OHT (1 μM), or MitoQ (1 μM) detected by acridine orange staining. Scale bar, 50 μm. F The mean intensity ratio of red fluorescence to green fluorescence in acridine orange staining (n = 3 per group). G Lysosomes staining in rat chondrocytes treated with RSL3 (0.25 μM), 4OHT (1 μM), or MitoQ (1 μM) detected by lyso-tracker red. Scale bar, 100 μm. H The mean fluorescence intensity in lyso-tracker red staining (n = 3 per group). I Flow cytometry analysis of ROS level of rat chondrocytes after RSL3 (0.25 μM), 4OHT (1 μM), or MitoQ (1 μM) treatment (n = 3 per group). J The merged images of Lyso-tracker red, Mito-tracker green, and Hoechst 33258 staining in rat chondrocytes treated with RSL3 (0.25 μM), 4OHT (1 μM), ScpI2 (5 μM), or CQ (20 μM). Scale bar, 50 μm. K The relative colocalization coefficients of mitochondria and lysosomes with Mito-tracker green and Lyso-tracker red staining (n = 6 per group). Data are expressed as means + SD. Unpaired two-tailed t tests, *P < 0.05.
