Overexpression of mimitin decreases caspase 3/7 activities. HepG2 cells were transfected either with pcDNA3 empty vector, or pcDNA3mim and then not-treated (control) or treated with TNF/CHX. Caspase activity was determined using Caspase-GloTM3/7 reagent. A. Graph presents changes in the relative changes in caspase 3/7 activity. In each case the value of caspase 3/7 activity after TNF/CHX treatment was divided by the value of basal activity of nonstimulated cells. Caspase activity in cells overexpressing mimitin was compared to cells transfected with an empty vector pcDNA3 (assumed as equal 1). The values represent the mean of 3 independent experiments ± SD with *p < 0.05. B. Western blot confirming overexpression of mimitin in cells transfected with pcDNA3mim. The blot was later stripped and reprobed with an α-tubulin antibody (lower panel) to ensure equal loading. RT-PCR results show transcript level for neomycin resistance gene (exogenous gene in pcDNA3) as a control of transfection efficiency and transcript level for elongation factor 2 (endogenous gene) as a control of RNA amount in tested samples.
Wegrzyn et al. BMC Cell Biology 2009 10:23 doi:10.1186/1471-2121-10-23