#Serum vst buffer plus

ER stress is caused by the accumulation of unfolded or malfolded proteins in the ER (Harding et al., 1999, Rao et al., 2004, Lindholm et al., 2006).

The endoplasmic reticulum (ER) plays key roles in the quality control of proteins via the folding and post-translational modification of newly synthesized proteins. These results suggest that the NGF-activated PI3-K/Akt signaling pathway plays a protective role against ER stress-mediated apoptosis via enhanced expression of GRP78 in PC12 cells. RT-PCR and Western blot analyses revealed that enhanced production of nuclear p50 ATF6, but not spliced XBP1, mainly contributed to the NGF-induced enhancement of GRP78 expression in 2DG-treated cells. In addition, versipelostatin (VST), a specific inhibitor of GRP78 expression, and small interfering RNA (siRNA) against GRP78 mRNA also decreased both the effects of NGF on 2DG-induced apoptosis and GRP78 expression.

#Serum vst buffer plus

When a specific inhibitor of phosphatidylinositol 3-kinase (PI3-K), LY294002, was added to 2DG plus NGF-treated cells, both the effects of NGF on 2DG-induced apoptosis and GRP78 expression were significantly diminished. Surprisingly, GRP78 expression was further up-regulated when NGF was added to 2DG-treated PC12 cells. We found that nerve growth factor (NGF) prevented 2DG-triggered ER stress-mediated apoptosis, but not the induction of GRP78 expression, in PC12 cells.

During ER stress, GRP78 expression is induced as part of the unfolded protein response (UPR). Glucose-regulated protein 78 (GRP78) is a molecular chaperone that acts within the ER. Inhibition of glycosylation caused by glucose depletion induces endoplasmic reticulum (ER) stress with subsequent apoptosis. The glucose analog 2-deoxy- d-glucose (2DG) depletes cells of glucose.