RNA-seq identified that ferroptosis may be involved in anti-osteoporosis effect of ACL under FAC exposure. (A) KEGG pathway enrichment analysis of all differentially expressed genes (DEGs) between ACL and ACL + FAC groups. (B) KEGG pathway enrichment analysis of down-regulated genes in ACL + FAC treated zebrafish larvae compared to ACL treated larvae. (C) Heatmap of 11 ferroptosis-related genes identified from DEGs. FAC represents the group treated with FAC alone, while ACL represents the group treated with both FAC and ACL. Each row represents one gene, and each column represents one sample. Red represents genes with high expression, blue represents genes with low expression. Darker colors indicate more significant differences. (D) Verification of mRNA expression level of ferroptosis-related genes in 5 dpf zebrafish larvae treated with 200 µg/mL FAC with or without 0.5 mg/L ACL. (E) Quantitative analysis of mRNA expression levels of three apoptosis genes, P53, casp3a and casp9. * means p < 0.05 between two indicated groups (n = 6). Data are displayed as mean ± SD.
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