Fig. 1: Accumulation of 24S-OHC esters activated the UPR signaling pathway and downregulated expression of ER chaperone proteins in SH-SY5Y cells.

a–c SH-SY5Y cells were pretreated with 5 μM F12511 for 15 min or with 100 μM Nec-1 (a) for 1 h and then exposed to 50 μM 24S-OHC for 6 h. Cells were also treated with 3 μM thapsigargin (Thapsi) for 3 h. a Whole-cell lysates were subjected to immunoblotting with appropriate antibodies as indicated. b The unspliced (XBP1u) and spliced (XBP1s) XBP1 mRNAs were analyzed by RT-PCR. c Whole-cell lysates were immunoblotted with antibodies specific for XBP1s or β-actin. d Cells were pretreated with 20 μM MG132 for 30 min and then exposed to 50 μM 24S-OHC or 3 μM thapsigargin for 6 h. Whole-cell lysates were immunoblotted with antibodies specific for ATF6 or β-actin. Asterisks denote nonspecific bands. e, f Cells were treated as in panel a. g, h Cells were treated as in panel d. e, g Whole-cell lysates were subjected to immunoblotting with appropriate antibodies as indicated. f, h Band intensities were quantified by densitometric scanning, relative intensity is shown. Mean ± SD n = 3, **P < 0.01, n.s. not significant