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. 2011 Apr 6;6(4):e18517. doi: 10.1371/journal.pone.0018517

Figure 5. Ufm1, Ufbp1and Ufl1 expressionis induced upon ER stress.

Figure 5

A mRNA expression level of Ufm1 (A), Ufbp1(B) or Ufl1 (C) after exposure of INS-1E cells to 25 µM CPA, 1 µM thapsigargin, 1 µg/ml brefeldin A, 5 µg/ml cycloheximide, 30 µM H2O2, 0.5 mMoleate and 0.5 mMpalmitate for 14 hours, measured via qPCR and normalized for GAPDH (Ufm1 and Ufbp1) or actin (Ufl1). Data are means±SEM, n = 4–6, *, p≤0.05; **, p≤0.01, paired student t-test, D UFM1 and UFBP1 protein expression is induced in cells exposed for 14 hours to CPA. Actin is shown as a control for protein loading, E UFM1-UFBP1 conjugates. Incubation of MIN6 or INS1 cells with ER stressors (25 µM CPA, 14h, INS1 cells; 1 µM thapsigargin (Tg), 1h, MIN6 cells) or proteasome inhibitor (100 µM MG115, 2h, INS1 cells) decreased conjugation, while incubation with a translational inhibitor (10 mg/l cycloheximide (CHX), 2h, MIN6 cells) increased the conjugate formation. * = UFM1−UFBP1 conjugate, 10 kDa = free UFM1.