Fig. 4

PSNPs treatment upregulates mitochondrial ROS generation to promote necroptosis in RAW 264.7 cells. (A) TNF-α quantification in RAW 264.7 cells treated with PSNPs for 4, 12, 24 h. (B) Total intracellular ROS levels in RAW 264.7 cells treated with 50 µg/mL PSNPs from 10 min to 1 h. (C) RAW 264.7 cells were treated with PSNPs with or without Mito-TEMPO or NAC for 30 min. The level of cytosolic ROS levels and mtROS were determined by dichlorodihydrofluorescein diacetate (DCFH-DA) staining and MitoSOX red staining, respectively. (D) Detection of late apoptotic/necroptotic cells in RAW 264.7 cells treated with or without Mito-TEMPO or NAC for 4 h. The results are presented as the mean ± S.D. n = 3, N.S., no significance. * P < 0.05, ** P < 0.01