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. 2017 Apr 15;58(6):1100–1113. doi: 10.1194/jlr.M074278

Fig. 6.

Fig. 6.

Palmitate, but not elaidate or oleate, induces UPR and inflammation markers in RAW264.7 macrophages. RAW264.7 macrophages were treated for 6 h with individual FAs (500 μM) as indicated. A: RT-PCR of Xbp1 processing showing unspliced (u) and spliced (s) Xbp1 mRNA with L32 as control. B: Immunoblot of UPR downstream effectors and UPR responsive proteins using regular gels (BIP, PERK, CHOP, and XBP1) and phospho-tag gels (IRE1α). C: Quantitative PCR expression of selected UPR markers. D: Immunoblot of cleaved caspase-3, an indicator of activated cell death by apoptosis. E: Immunoblot of ER-associated protein degradation markers. F: Quantitative PCR expression of selected UPR and inflammation marker genes after treatment with the trans FAs, trans-vaccenic acid, palmitelaidic acid, CLA 9Z,11E, CLA 10E,12Z, and palmitate. G: Immunoblot of CHOP and XBP1 after treatment with different trans FAs and palmitate. H: Quantitative PCR expression of selected UPR and inflammation marker genes after treatment with the saturated FAs, laurate, myristate, palmitate, and stearate. I: Immunoblot of CHOP and XBP1 after treatment with different saturated FAs. mRNA expression was normalized against 36b4. Error bars represent SD. Asterisks indicate statistically significant relative to vehicle control (*P < 0.05).