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Figure 4


Fig. 4. FFA activate IRE1 and ATF6 signaling. INS-1E cells were cultured in the presence or absence of oleate (O, gray diamonds), palmitate (P, black squares) or oleate plus palmitate (OP, white circles) at a glucose concentration of 11 mM. The mRNA expression of BiP, XBP1s and XBP1t following a 6- to 48-hour FFA exposure was analyzed by real-time PCR, normalized for the expression level of the housekeeping gene GAPDH and expressed as fold induction. The results represent means ± s.e.m. of 6-11 independent experiments. For the measurement of UPR luciferase reporter activity, INS-1E cells were co-transfected with the UPR reporter, which is responsive to ATF6 and XBP1s, and with the internal control pRL-CMV, encoding Renilla luciferase. After overnight transfection, the cells were exposed for 24 hours to FFA. The firefly results were normalized for Renilla luciferase activity and are means ± s.e.m. of ten independent experiments. *P<0.05, **P<0.01, ***P<0.001 vs control (C).





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