Fig. 6: Regulation of GSK-3β and PERK by CO sustain nuclear TFEB levels, leading to suppressed amyloidogenesis.
From: CO confers neuroprotection via activating the PERK-calcineurin pathway and inhibiting necroptosis
a–d SH-SY5Y cells were treated with 20 μM CORM-A1 after the cells were transfected with CA-GSK-3β or treated with 1 μM GSK2606414. Nuclear and cytoplasmic fractions extracted from SH-SY5Y cells were analyzed by western blotting using TFEB antibody (left), and quantification of nuclear TFEB and cytosol TFEB was analyzed (right) (a). The levels of p-GS, GS, p-eIF2α, and eIF2α expression were assessed by western blotting (b). Quantification of p-GS and p-eIF2α (c). The protein expression of autophagy protein LC3-I/-II and lysosomal protein LAMP1 was detected by western blotting (left panel) (d), and quantification of LC3-II conversion and LAMP1 is shown in the right panels. e SH-SY5Y cells were transfected with APP-Swe/Ind and then treated with 20 μM CORM-A1 for 12 h in the presence or absence of 1 μM GSK2606414. The levels of APP, LAMP1, LC3-I/-II, p-eIF2α, and eIF2α expression were detected by western blotting. f SH-SY5Y cells were co-transfected with APP-Swe/Ind and siTfeb for 36 h and were treated with 20 μM CORM-A1 for 12 h. The levels of APP, LAMP1, LC3-I/-II, and TFEB were analyzed by western blotting. g SH-SY5Y cells were treated with 20 μM CORM-A1 after the cells were co-transfected with CA-GSK-3β and APP-Swe/Ind or treated with 1 μM GSK2606414. The protein expression of FL-APP was determined by western blotting (left), and quantification of FL-APP was analyzed (right). Data were expressed as mean ± SD; *p < 0.05, **p < 0.01, ***p < 0.001, and NS not significant.
