Fig. 6: Mitochondrial damage by 7-ketocholesterol induces dysregulation of the TNF signaling pathway in macrophages.

KEGG pathway enrichment analysis of differentially expressed genes between 7K-Cho-treated (a) or DOX-treated (b) and vehicle-treated BMDMs for 6 h. Enrichment analysis was performed using Fisher’s exact test. In order to control the calculation of false positives, BH method was provided to correct the P value (adjust P). n = 3 biological samples per group. c THP-1 cell luciferase reporter gene assays. THP-1 cells were co-transfected with the pTNF-α promoter luc plasmid, pNF-κb luc plasmid, pSTAT3 luc plasmid, or pRL-SV40 plasmid. After 24 h, the cells were harvested and cultured with 1% FBS medium containing vehicle, DOX (10 µg/ml), cholesterol (10 µg/ml), or 7K-Cho (10 µg/ml) medium for another 24 h. Cells were stimulated with LPS (100 ng/ml), TNF-α (20 ng/ml), or IL-6 (10 ng/ml) for 6 h, and cells were lysed to detect firefly luciferase and Renilla luciferase. n = 3 independent experiments. d Quantification of ex vivo phagocytosis in ApoA1- and TNF-α-inhibitor (CC5013)-treated TAMs. TAMs isolated from intracranial GL261-bearing mice were cultured with vehicle, ApoA1 (10 µg/ml), or CC5013 (20 nM) medium for 24 h; or TNF-α (20 ng/ml) for 6 h. n = 3 biological samples per group. Statistical significance was determined using the unpaired t test (Two-tailed) in c, d. All data are the mean ± SD. Source data are provided in the Source Data file.