Fig. 2

Rapamycin treatment limits caspase-3 activation to promote cell survival during CoCl₂ exposure. a Rapamycin treatment reduces initiator caspase-9 activation. HT22 cells were untreated or treated with 250 µM CoCl₂, with and without 500 nM rapamycin for 24 h. Cells were lysed to obtain cytosolic protein fractions and cleaved caspase-9 protein expression was analyzed by western blot. Left is the average relative protein expression of each group when normalized to total caspase-9 protein expression. Right is a representative image of at least three separate blots showing caspase-9 protein expression. ^###p < 0.001 versus control; ***p < 0.001 versus CoCl₂ alone. b Rapamycin treatment reduces executioner caspase-3 activation. Cells were treated, and cytosolic protein fractions obtained and analyzed as in A. Left is the average relative protein expression of cleaved caspase-3 when normalized to total caspase-3 protein expression. Right is a representative image of caspase-3 protein expression from at least three separate blots. ^###p < 0.001 versus control; ***p < 0.001 versus CoCl₂ alone. c Rapamycin does not affect CoCl₂-induced cytochrome C release. Cells were treated, and cytosolic protein fractions obtained and analyzed as in A. Left is the average relative protein expression of cytochrome C when normalized to β-actin protein expression. Right is a representative image of cytochrome C protein expression from at least three separate blots. ^##p < 0.01 versus control