Fig. 5: TFEB nuclear export is prevented by CO-induced GSK-3β inhibition.
From: CO confers neuroprotection via activating the PERK-calcineurin pathway and inhibiting necroptosis
a To assess the effect of CO on inhibition of TFEB nuclear export, SH-SY5Y cells transiently expressing TFEB-GFP were starved for amino acids for 2 h and then refed with complete DMEM for 2 h in the presence or absence of 20 μM CORM-A1 and the nuclear export inhibitor LMB (20 nM) and analyzed by confocal microscopy. b SH-SY5Y cells were co-transfected with TFEB-GFP, CA-GSK-3β, or KD-GSK-3β. The transfected cells were treated with 20 μM CORM-A1 for 2 h after cells were refed with complete DMEM. Fluorescence images of SH-SY5Y cells were obtained by confocal microscopy. S (Starved), S + R (Starved + Refed), S + R + C (Starved + Refed + CORM-A1). c, d SH-SY5Y cells were placed in HBSS medium for 2 h and then refed with complete DMEM medium for 2 h in the presence or absence of 20 μM CORM-A1. Images of TFEB-GFP were detected by confocal microscopy in SH-SY5Y cells transfected with siRNA against GSK-3β (c) or treated with LiCl (d). The percentage of cells showing nuclear TFEB localization was calculated. n > 20 cells per condition. Data were expressed as mean ± SD; ***p < 0.001 and NS not significant.
