Fig. 4

EgCF promoted transcription and protein stability of KDM5B. The expressions of histone demethylase gene (A) and differentially expressed KDM5B (B) were obtained from RNA-seq data of mouse peritoneal macrophages treated with LPS and EgCF. Mouse peritoneal macrophages were treated with EgCF (50% v/v) and LPS (1 μg/ml) for 6 h, and mRNA expression levels of KDM5B, KDM6B, and KDM7A (C) were detected by qRT-PCR. RAW 264.7 cells were treated with EgCF (50% v/v) and LPS (1 μg/ml) for 6 h, and mRNA levels of KDM5B were measured by qRT-PCR (D). Protein levels of KDM5B were measured by Western blotting (E). THP-1 cells were treated with EgCF (50% v/v) and LPS (1 μg/ml) for 6 h. mRNA levels of KDM5B were measured by qRT-PCR (F), and protein levels of KDM5B were measured by Western blotting (G). Protein levels of KDM5B were measured in THP-1 cells stimulated with EgCF (50% v/v) and LPS (1 μg/ml) for 15, 30, 60, 120, and 240 min (H). RAW 264.7 cells were treated with cycloheximide (CHX, 50 μg/ml) in combination with EgCF and LPS for 2, 4, and 6 h. Protein levels of KDM5B were measured by Western blotting (I). RAW 264.7 cells were treated with the SUMOylation inhibitor TAK-981 (0.5 µM and 1.0 µM) in combination with EgCF and LPS for 6 h, and KDM5B protein levels were measured by Western blotting (J). C, D, F Data are presented as the mean + SEM of three independent experiments and compared using the one-way ANOVA and Tukey test. Asterisks indicate a significant difference at *P < 0.05 and **P < 0.01