The involvement of survival signaling pathways in rubella-virus induced apoptosis

Cooray, Samantha; Jin, Li; Best, Jennifer M

RV induces apoptosis in RK13 cells with characteristic morphological and biochemical features [6, 8, 9] . The XTT assay was used to examine the effect of RV infection and LY29002 and U0126 treatment on cellular metabolism over time. XTT is a tetrazolium salt, which is cleaved by the succinate dehydrogenase system of mitochondria in Kinase phosphorylation during RV infection Figure 1 Kinase phosphorylation during RV infection. Serum-starved RK13 cells were mock infected or infected with RV at an m.o.i. of 4 PFU/cell. At indicated time points cell lysates were collected and proteins (30 µg/lane) were separated by SDS-PAGE, and analysed by Western blotting using phospho-specific antibodies. Blots were also probed with anti-tubulin antibody to demonstrate equal loading. A -Total and phosphorylated Akt and ERK (24-96 hours p.i.). B -Total and phosphorylated Akt, ERK, and p70S6K, and phosphorylated GSK-3β and c-myc. The data were consistently repeated in two independent experiments. metabolically active cells, to yield a soluble orange formazan product. A decrease in the intensity of formazan was used to monitor changes in cellular metabolism and cell viability in RV-infected cells by spectroscopy.