Fig. 4.

p62/Nrf2/ARE axis was involved in autophagy blockage-mediated aggregation of macrophage pyroptosis. In the presence of ox-LDL (75 μg/mL), cells were stimulated by a Nrf2 agonist (tBHQ) or inhibitor (ML385) respectively. Next, (a, b) HO-1 and Nrf2 protein level were measured by immunoblotting. Cells were pretreated with CQ for 24 h and then tBHQ (20 μM) or ML385 (5 μM) were added to the cells for an additional 24 h in the present of ox-LDL. (c, d, e) Relative protein and mRNA levels of p62, LC3II/I, p-casp1, GSDMD, HO-1 and Nrf2 determined by immunoblotting and PCR. (f) Cell viability and (g) LDH release were detected in the four groups. (h, i) IL-1β and IL-18 secretion were assessed by ELISA. (j) FCM was used to test the radio of stained cells. (k) A network of related proteins and their co-interaction was analyzed using STRING and cytoscape. The thickness of the line presents the degree of their relationships. Values are expressed as the mean ± SEM (n = 3) of independent experiments at least 3 repeated measurements. ^*p < 0.05 and ^**p < 0.01 different from the control group, ^#p < 0.05 and ^##p < 0.01 different from the ox-LDL-treated group, and ^△p < 0.05 and ^△△p < 0.01 different from the ox-LDL+CQ-treated group