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. 2017 Sep 27;292(49):19976–19986. doi: 10.1074/jbc.M117.801936

Figure 6.

Figure 6.

Role of N-terminal residues of DESAT1 in the regulation of degradation of DESAT1. A, N-terminal amino acid sequences of wild-type and mutants of DESAT1. S2 cells expressing DESAT1 (N4A/Q6A)-FLAGC, DESAT1 (P2A/P3A)-FLAGC, DESAT1 (P2A)-FLAGC, and DEAT1 (P3A)-FLAGC were treated with C18:1 (100 μm) for 6 h (B) or DESAT1 inhibitor 37c (1 μm) for 16 h (C), and the amounts of DESAT1 and α-tubulin protein were detected with anti-DESAT1 antibody and anti-α-tubulin antibody, respectively. S2 cells expressing DESAT1 (P2A/P3A)-FLAGC were treated with cycloheximide (100 μg/ml) for 0, 2, and 4 h, and the amounts of DESAT1 and α-tubulin protein were detected with specific antibodies (D and E). Band intensities were determined by ImageJ software, and levels of DESAT1 proteins are shown relative to the amount of DESAT1 protein in vehicle-treated cells (B and C) or cells not treated with cycloheximide (E). The values for DESAT1-FLAGC (Fig. 3, D and F) are shown for comparison (B and C). Filled arrowhead, DESAT1 (P2A/P3A)-FLAGC; open arrowhead, endogenous DESAT1. Mean ± S.D. (n = 3). *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s., not significant.