Figure 5.

E₂ promotes trained immunity in macrophage cell lines J774 and RAW264.7. (A,B) The effect of E₂ on both J774 and RAW264.7 cell viability for 24 h was tested with CCK-8 assay kit. (C) In vitro trained immunity model for J774 and RAW264.7 cell line. (D) The mRNA levels of TNFα and IL-1β in J774 were detected by qPCR to determine the trained immunity effect with or without E₂. (E) The mRNA levels of TNFα and IL-1β in RAW264.7 were detected by qPCR to determine the trained immunity effect with or without E₂ (n ≥ 3/group). ^#p < 0.05, ^###p < 0.001, paired Student's t-test comparing β-glucan + LPS group and LPS group. *p < 0.05, ***p < 0.001, paired Student's t-test comparing with control group. ∧p < 0.05, ∧∧p < 0.01, and ∧∧∧p < 0.001, paired Student's t-test comparing between β-glucan + LPS groups with or without E₂.