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. 2021 Sep 15;22(18):9952. doi: 10.3390/ijms22189952

Figure 3.

Figure 3

Effect of the acute glucose shift-induced generation of ROS on the activation of the NLRP3 inflammasome in THP-1 cells. Before any glucose shift, cells were pretreated with a pharmacological antioxidant, NAC (10 mM) for 1 h and then exposed to a glucose shift for 24 h. (A) The intracellular ROS level was measured by flow cytometry using CM-H2DCFDA. (B) Mean fluorescence intensity of the levels of ROS is presented as a percentage of the NG control. (C) Cell lysates were subjected to western blotting using antibodies against NLRP3, caspase-1, and IL-1β. (D–F) The protein expression levels of NLRP3, caspase-1, and IL-1β were quantified using ImageJ. (G) The level of IL-1β in the culture supernatant was determined by ELISA. Data are presented as the mean ± SEM from at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.