Activation of the Mitochondrial Apoptotic Signaling Platform during Rubella Virus Infection

Claus, Claudia; Manssen, Lena; Hübner, Denise; Roßmark, Sarah; Bothe, Viktoria; Petzold, Alice; Große, Claudia; Reins, Mareen; Mankertz, Annette; Frey, Teryl K.; Liebert, Uwe G.

For subsequent experiments z-VAD-fmk was used at 12.5 μM such that the lowest possible effective concentration was used for its application at 24 hpi. While PFTμ and z-VAD-fmk were both applied at 24 hpi, 2 hpi was the selected application time point of NIM811, as this time point was slightly more effective than the one at 24 hpi. Figure 1B indicates that viral titer was not affected by application of these inhibitors, which otherwise could also result in a reduction of RV-associated 3 Figure 1 . Analysis of selected pharmacological inhibitors (z-VAD-fmk (12.5 µM if not otherwise indicated), PFTµ and α (12.5 µM), and NIM811 (2 µM)) with respect to RV replication and cytopathic effect (CPE) induction. (A) Effect of different application time points on the presence of floaters in the supernatant of RV-infected Vero cells at 3 dpi. The respective SC (DMSO)-treated sample was set at 100%; (B) RV titer was determined by plaque assay for supernatants collected at 3 dpi after drug application at the time point with maximal efficacy (NIM811 at 2 hpi, z-VAD-FMK and PFTµ at 24 hpi). PFT α was added at 2 hpi. In comparison to the SC, changes in viral titer were not significant; (C) Characterization of the presence of an apoptotic laddering in the floater population collected from the supernatant of the SC-and NIM811 (2 hpi)-and PFTµ (24 hpi)-treated and RV-infected Vero cells.