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. 2021 Jan 26;22(3):1194. doi: 10.3390/ijms22031194

Figure 2.

Figure 2

Inhibitory effect of KMU-1170 on LPS-induced upregulation of proinflammatory cytokines in THP-1 cells. Cells were differentiated into macrophages for 24 h using PMA (100 nM). And then the cells were treated with LPS (1 μg/mL) for 6 h after pretreatment with different doses of KMU-1170 (0.1, 0.5, and 1 μM) for 1 h. (A) Whole cell lysates were isolated and used to measure the protein expression levels of pro-IL-1β, TNF-α, IL-6, and β-actin by Western blot analysis. (B) Image-J software was used to analyze the relative optical density of the pro-IL-1β, TNF-α, and IL-6 band, respectively. (C) Total RNA was extracted and used to determine the mRNA expression levels of IL-1β, TNF-α, and IL-6 using RT-PCR. (D) Image-J software was used to analyze the relative optical density of the IL-1β, TNF-α, and IL-6 band, respectively. (E–G) Total RNA was extracted and used to determine the mRNA expression levels of IL-1β, TNF-α, and IL-6 using real time PCR. † p < 0.05, * p < 0.01, and # p < 0.001 compared to LPS alone.