Figure 7
Drp1 cells are resistant to apoptotic signals. (a) Growth curves. In all, 105 Vector and Drp1 cells were seeded onto 100-mm tissue culture plates in growth medium containing 10% fetal bovine serum and allowed to attach overnight. The following day (day 0), cells were counted and fresh growth medium either lacking or containing 4-HT was then added. Viable cell counts were then performed daily using a Vi-Cell analyzer. The results shown represent the mean of triplicate determinations±1 S.E. (b) Apoptosis profiles. Vector and Drp1 cells were seeded onto 100-mm plates in growth medium and allowed to achieve ∼80% confluency at which point (day 0) the total number of viable cells was determined from triplicate cultures. The remaining monolayers were then maintained for the duration of the experiment in serum-free medium either lacking or containing 4-HT, and the number of viable cells was determined daily thereafter. (c) Delayed cytochrome c release from Drp1 cells. Cells were initially plated and grown as described in panel (b) and then deprived of serum upon reaching ∼80% confluency. At the indicated time points, the cells were harvested and fractionated into cytosolic and mitochondrial fractions. Immunoblotting was then performed to allow for detection of cytochrome c release from the mitochondria. Loading controls consisted of β-actin for cytosolic fractions and Ponceau Red staining of the low molecular weight region of the membrane corresponding to the region of cytochrome c migration
