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. 2010 Apr 12;107(17):7892–7897. doi: 10.1073/pnas.1003585107

Fig. 4.

Fig. 4.

Fatty acid-stimulated posttranslational regulation of PNPLA3. (A) Dose–response, (B) time course, and (C) effect of fatty acid saturation and chain length on PNPLA3 protein mass. (A) HuH-7 cells stably expressing PNPLA3-V5 were treated with oleate for 8 h. Immunoblotting of the cell lysates was performed using anti-V5 and calnexin antibodies. (B) Cells were treated with 400 μM oleate for the indicated times before immunoblotting. RNA was extracted from duplicate sets of cells and the abundance of PNPLA3 mRNA was assayed using real-time PCR. (C) Cells transfected with 0.5 μg of pTK-Insig1-myc (34) were treated for 6 h with vehicle or 400 μM palmitate (C16:0), oleate (C18:1), linoleic acid (C18:2), linolenic acid (C18:3), arachidonic acid (C20:4), and eicosapentaenoic acid (C20:5), respectively. Cell lysates were subjected to immunoblotting. (D) Cultured cells were treated with 400 μM oleate plus DMSO or 5 μM triacsin C for the indicated times before immunoblotting.