Inhibiting β-Oxidation Attenuates ER Stress Induced by Overexpression of a Constitutively Misfolded Protein
(A) Primary hepatocytes were infected with Ad-CMV-eGFP or Ad-TetOn-NHK at an MOI of 1. NHK expression was induced with 500 ng/mL doxycycline (Dox) in the presence or absence of 25 μg/mL ET. mRNA expression of UPR markers was measured by qRT-PCR after 8 h of treatment.
(B) Primary hepatocytes infected with Ad-TetOn-NHK and treated with Dox and/or ET for 8 h were fixed in 2.5% glutaraldehyde for TEM. Orange arrowheads in insets denote dysmorphic ER. Images from which these insets were taken, along with scale bars, are in Figure S4.
(C) Images were scored as described in Figure 2; n = 10 cell images per group.
(D) Primary hepatocytes infected with Ad-TetOn-NHK were treated with Dox and ET as in (A) for 4 or 8 h in the presence or absence of 5 μM MG-132. NHK expression was analyzed by immunoblot. Loading control was calnexin.
See also Figure S4.