Skip to main content
. 2021 Nov 10;12:737065. doi: 10.3389/fimmu.2021.737065

Figure 2.

Figure 2

DMF decreased the expression of inflammasome complex proteins, prevented caspase-1 activity and ASC speck formation. N9 microglia were pretreated with DMF (10 µM) for 1h, primed with UP-LPS (1000 ng/ml) for 4 hours, and followed by ATP (5 mM) for 1 hour. Pretreatment with DMF led to decrease expression of NLRP3, inhibited cleavage of pro-caspase 1(p45) into caspase-1(p20), and prevented caspase-1 activity and ASC speck formation in microglia. (A–C) The expression of pro-caspase-1 and its cleavage were evaluated with Western blot. (D) Caspase-1 activity was measured with a luminometric assay. (E) mRNA expression of NLRP3 was measured with qRT-PCR. (F, G). Protein expression of NLRP3 was detected by Western blot. (H, I) ASC speck formation was demonstrated by the IF method (Blue staining – Hoechst, Red staining – ASC). Data are presented as mean ± S.E.M, n = 5. *p < 0.05, **p < 0.01 compared to control and #p < 0.05, ##p < 0.01 compared to LPS and ATP induced cells.