Induction of a Feed Forward Pro-Apoptotic Mechanistic Loop by Nitric Oxide in a Human Breast Cancer Model
Figure 3
Immunoblot analysis of the dephosphorylation of pFOXO1 at serine 256, in MDA-MB-468 cell lysates, over time following treatment with 1 mM DETA-NONOate [a, left panel]. Immunoblot analysis of the dephosphorylation of pFOXO1 at serine 256 in MCF-7 cell lysates at 24 h post-treatment with 1 mM DETA-NONOate [a right panel].
Immunoblot analysis of dephosphorylation of pFOXO1 upon DETA-NONOate treatment in either scrambled controls or MDA-MB-468 cells silenced for the catalytic subunit of PP2A [PP2A siRNA] [b]. Immunoblot analysis of FOXO1 in MDA-MB-468 nuclear fractions at 24 h post-treatment with 1 mM DETA-NONOate [c ]. Silencing of FOXO1 in MDA-MB-468 cells utilizing siRNA against human FOXO1 [d left panel] qPCR analysis to determine the fold change in mRNA expression of FOXO1 in MDA-MB-468 transfected with either scrambled controls [SiScramble] or siRNA against the human FOXO1 gene [siFOXO] [d, right panel]. Annexin V binding assay at 24 h, in MDA-MB-468 cells either transfected with scrambled controls [siScramble] or siRNA against the human FOXO1 gene, following treatment with 1 mM DETA-NONOate [1 mM DETA] [e]. Caspase-3 activity in MDA-MB-468 at 24 h following treatment with 1 mM DETA-NONOate in either scrambled controls [siScramble] or silenced for human FOXO1 gene [siFOXO] [f]. Results are expressed as the means from three independent experiments performed in duplicate; bars, SE wherever applicable. Western blot images are representative of three different experiments. GAPDH and Lamin A are probed as equal loading controls for cytosolic and nuclear nuclear fractions respectively.