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. 2011 Oct 14;286(48):41862–41870. doi: 10.1074/jbc.M111.304956

FIGURE 6.

FIGURE 6.

The human TAP1 mRNA is a direct target of miR-346. A, the predicted seeding of miR-346 to the 3′-UTR of the human TAP1 mRNA. The human TAP1 mRNA 3′-UTR and the predicted binding site of miR-346 are shown with the 6-mer canonical seeding site highlighted in gray. AU elements in the vicinity of the predicted miR-346 binding are indicated with a rectangle. B, ER stress reduces TAP1 mRNA levels in Calu-3 and HeLa cells. Cells were treated with different ER stressors, TM, proteasome inhibition (PI), thapsigargin (TG), and brefeldin A (BFA), for 12 h, and relative TAP1 mRNA levels were measured. Results are plotted relative to untreated control TAP1 mRNA level (n = 4). C, ER stress reduces TAP1 protein levels. The effects of ER stress (TM) were tested on human TAP1 protein levels in HeLa cells. Cells were treated with TM (5 μg/ml) for the time periods specified, and 30 μg of total protein lysate was analyzed by SDS-PAGE and Western blot analysis for TAP1 expression. Actin was detected as a loading control, and densitometry was performed using ImageJ (National Institutes of Health). The percentage values represent the percentage of untreated control (100%). D, ER stress has no effect on mouse TAP1 mRNA levels. Mouse embryonic fibroblasts (Xbp1+/+ and Xbp1−/−) were treated with TM (5 μg/ml) for 18 h to induce ER stress. The relative mouse TAP1 mRNA levels were determined by qRT-PCR using mouse TAP1-specific primer sets. Results are plotted relative to untreated control (n = 3).