Assessment of cell viability of Bhk-21 cells at low and high concentrations of Tunicamycin after transfection of Bcl-2 ER localized proteins

It is known that the B-cell lymphoma (Bcl-2) family of proteins has the ability to signal for apoptosis under stressful conditions to the cell, as well as to arrest apoptosis. The ability for Bcl-2 proteins to do this is due to their homology and their interaction with proteins such as the inositol 1,4,5-triphosphate (IP3) receptor, a calcium channel in the endoplasmic reticulum (ER). Blocking the IP3 receptor with 2- Aminoethoxydiphenyl borate (2-APB) has proven to be sufficient to block apoptosis at low levels of ER stress. In this study, Bcl-2 was used in an attempt to block apoptosis at low levels of ER stress. ER localized Bcl-2 (ER-Bcl-2-GFP) plasmid constructs were used to transfect Baby Hamster Kidney (BHK21) cells then exposed t plow and high levels of ER stress in an attempt to replicate the rescue 2- APB had on the temperature sensitive hamster firbroblasts (tsBN7). It was shown that the over expression of ER-Bcl-2 was able to sufficiently rescue tsBN7 cells from apoptosis under low levels of ER stress through temperature shifts. Also, it was proven that BHK21 cells were sufficiently rescued by 2- APB (50μM) under low levels (30nM) of Tunicamycin induced ER stress. Though, after transfections, and 36 hour exposure to low (20nM-30nM) and high (200nM) levels of Tunicamycin induced apoptosis, ER-Bcl-2 was not able to significantly rescue BHK21 cells. Albiet, there was a trend in some of the trials, but no replicates allow for significanc