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. Author manuscript; available in PMC: 2021 Nov 1.
Published in final edited form as: J Biochem Mol Toxicol. 2020 Jun 24;34(11):e22561. doi: 10.1002/jbt.22561

FIGURE 4.

FIGURE 4

PFOA-mediated ER stress precedes oxidative stress. A, Immunoblot analysis for Atf4 and Chop expression in 266-6 cells following 1 hr pretreatment with 5µM 4-phenylbutyrate (4PB), 1mM N-acetyl-cysteine (NAC), or 0.4mM 4,5-Dihydroxy-1,3-benzenedisulfonic acid disodium salt monohydrate (Tiron) prior to treatment with DMSO (−), 50 µg/mL PFOA or 10µM H2O2 for 6 hrs. Gapdh expression is included as a loading control. Shown is a representative experiment (n = 4). Noted are the phosphorylated forms of Atf4 detected by the Atf4 antibody while NS indicates recognition of a non-specific band. B-D, mRNA expression of Atf4, Chop and Sod1 was determined by qRT-PCR analysis following 1 hr pretreatment of 266-6 cells with 5µM 4-phenylbutyrate (4PB), 1 mM N-acetyl cysteine (NAC), or 0.4mM 4,5-Dihydroxy-1,3-benzenedisulfonic acid disodium salt monohydrate (Tiron) followed by treatment with DMSO (CTL), 50 µg/mL PFOA or 10µM H2O2 for 6 hrs. Quantitation is expressed as fold-induction ± std dev of assays performed in triplicate with DMSO (CTL) levels set to 1.0 from a representative experiment (n = 3). * indicates significantly different from DMSO (CTL) p < 0.05; # indicates significantly different from respective PFOA or TG values p < 0.05.