Fig. 3

Elevated inflammatory cytokines in the spinal dorsal horn of ApoE4-TR mice. (A) Experimental scheme of ELISA and RT-qPCR in mice. (B) Quantification of cytokine levels (IL-6, IL-1β, and TNF-α) in the spinal dorsal horns of the injured side of ApoE3-TR and ApoE4-TR mice by ELISA 14 days after SNI. n = 3 biologically independent samples. (C) RT-qPCR analysis of IL-6, IL-1β, and TNF-α in the dorsal horn of ApoE3-TR and ApoE4-TR mice 14 days after SNI. n = 3 biologically independent samples. (D) Representative images of the cultured primary astrocytes stained by anti-GFAP (red) and DAPI (blue). Scale bar, 20 μm. (E) Quantification of GFAP⁺ cells. Each data point represents the mean percentage of GFAP⁺ cells per well, with n = 5 wells per group. (F) Experimental scheme of ELISA and RT-qPCR in cultured astrocytes. (G) Quantification of cytokines released from ApoE3 and ApoE4 astrocytes by ELISA at 3 min and 12 h after medium replacement. n = 3 biologically independent samples. (H) RT-qPCR analysis of IL-6, IL-1β, and TNF-α in ApoE3 and ApoE4 astrocytes after CYM treatment. n = 3 biologically independent samples. All data are expressed as mean ± SEM. Statistical comparisons were conducted with one-way ANOVA followed by Tukey’s post hoc test (B, C); and unpaired Student’s t-test (E, G, H). *P < 0.05, and **P < 0.01