Fig. 2: PERK and IRE1-XBP1 axes were not involved in 24S-OHC-induced cell death in SH-SY5Y cells.

a SH-SY5Y cells were treated as in Fig. 1a. Whole-cell lysates were immunoblotted with antibodies specific for ATF4, CHOP, or β-actin. b, c Cells were transfected with CHOP (siCHOP #1-#3) or negative control (NC) siRNA oligo for 24 h. b Whole-cell lysates were immunoblotted with antibodies specific for CHOP or β-actin. Relative expression levels of CHOP are shown. c The cells were exposed to 50 μM 24S-OHC for 24 h. Cell viability was measured by LDH assay. **P < 0.01, when compared with cells treated with vehicle or among 24S-OHC-treated groups. d Cells were pretreated with 5 μM GSK2606414 for 30 min then exposed to 50 μM 24S-OHC or 3 μM thapsigargin for 6 h. Whole-cell lysates were immunoblotted with antibodies specific for PERK, IRE1α, or β-actin. e Cells were pretreated with 5 or 10 μM GSK2606414 for 30 min then exposed to 50 μM 24S-OHC for 24 h. Cell viability was measured by WST-8 assay. **P < 0.01. f Cells were pretreated with 5 μM F12511 for 15 min or with 1–10 μM 4μ8 C for 1 h then exposed to 50 μM 24S-OHC for 6 h or 1 μM thapsigargin for 3 h, respectively. XBP1 mRNAs were analyzed by RT-PCR. g Cells were pretreated with 3 or 10 μM 4μ8 C for 1 h and then exposed to 50 μM 24S-OHC for 24 h. Cell viability was measured by WST-8 assay. **P < 0.01