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. 2022 Feb 16;2022:8603427. doi: 10.1155/2022/8603427

Figure 5.

Figure 5

Rheb activates the NLRP3 inflammasome to enhance neuronal autophagy and apoptosis. (a) Rheb and NLRP3 were coexpressed as determined using the MEM tool. (b) NLRP3 expression was estimated by RT-qPCR. (c) NLRP3 expression in the OGD/R-exposed cell model was measured by RT-qPCR. (d) oe-NLRP3 neuron transfection efficiency was quantitated by RT-qPCR. (e) Rheb and NLRP3 expression in oe-Rheb+oe-NC cotransfected neurons exposed to OGD/R conditions as measured by RT-qPCR. (f, g) The secretion levels of IL-18 (f) and IL-1β (g) from neurons cotransfected with oe-Rheb+oe-NC and exposed to OGD/R conditions were measured by ELISA. (h) The activity of neurons was determined using a CCK-8 kit. (i) Expressions of autophagy-linked proteins (Beclin-1, LC3 II/I, LAMP-1, and P62) and NLRP3 inflammasome activation-related proteins as measured by western blot analysis. (j) Protein expressions of total caspase3 and cleaved-caspase3 in neurons were determined by western blot analysis. (k) The number of TUNEL+ neurons was determined by TUNEL staining. ∗p < 0.05 vs. control or oe-NC-transfected neurons. #p < 0.05 vs. oe-Rheb+oe-NC-cotransfected neurons (n = 3).