Figure 2. Administration of acALY18 to fibroblasts does not induce ER stress; however, ER stress induces acALY18. (A) A time course measuring the release of acALY18 from TRPC1 when human fibroblasts are treated with 25 nM thapsigargin. Results (top left) show that acALY18 expression precedes GRP78 expression (top right). Western analysis shows that acALY18 does not induce GRP78 in human fibroblasts treated with 25 nM thapsigargin (Tg), acALY18 (5 ng/ml), or mock treatment, for 24 h (bottom). NT, no treatment. (B) Normal human fibroblasts were treated with 25 nM thapsigargin, acALY18 (5 ng/ml), or mock treatment. Western analysis shows phosphorylation of eIF2α (p-eIF2α) after 1 h thapsigargin (Tg) treatment, but not acALY18 or mock treatment (left). Densitometry of the results is plotted (right) as the percentage p-eIF2α of total eIF2α. The expression of IRE1α after 24 h is only detectable in the thapsigargin treated cells (bottom).