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. 2020 Jun 30;17(13):4716. doi: 10.3390/ijerph17134716

Figure 3.

Figure 3

GR extract attenuated LPS-induced inflammatory responses and ER stress with the induction of antioxidant gene expression in mouse bone-marrow-derived macrophages (BMDMs). BMDMs were pretreated with GRE (50–250 μg/mL) for 48 h in FBS-containing completed media and then starved in DMEM for 12–18 h, followed by LPS stimulation (100 ng/mL) for 2 h (A,D) or 0.5, 2, and 4 h (B,C), in the presence or absence of GRE. (A) TNFα gene expression by qPCR. (B) TNFα and IL-1β gene expression by qPCR. (C) TNFα cytokine production by ELISA. (D) Protein extracts from LPS-treated BMDMs were immunoblotted with antibodies targeting p-JNK, p-ERK, p-p38, t-ERK, or β-actin (loading control); BMDMs were pretreated with GRE (100 μg/mL) for 48 h in FBS-containing completed media and then starved in DMEM, followed by LPS stimulation (100 ng/mL) for 4 h in the presence or absence of GRE. (E) GPx, SOD1, SOD2, and HO-1 gene expression by qPCR. All values are presented as the mean ± SEM (n = 3–4/group for each experiment). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; compared with LPS-treated group vs. LPS + GRE-treated group by one-way ANOVA with Bonferroni’s comparison test. +, treatment; −, non-treatment.