Fig. 5

Inactivating the HML-10(DAP3) RNA induces DAP3 expression and apoptosis in HeLa cells. a Target regions of sequence-specific ASOs are indicated. ASOs 1-4 are in antisense orientation to the retroviral transcript and in sense orientation to the DAP3 transcript. The ASO designated as Upstream served as control. b Cells were transfected with 25 or 50 nM of the indicated ASOs. At 24 h after transfection, expression levels of HML-10(DAP3) (left) and DAP3 mRNA (right) were determined by qRT-PCR. Bars show mean ± SEM of three independent experiments. RNA levels were normalized to GAPDH and levels of non-transfected cells were set to 1. *P-value ≤ 0.05, Student’s t-Test against Mock. c Cells were transfected with the indicated ASOs at 50 nM, after 24 h stimulated with 1000 U/mL IFNγ or 100 ng/mL TNFα, or left unstimulated. After additional 24 h, Trypan Blue exclusion as indicator of dead cells (left), MTS cell viability assays (center) or light microscopic analysis (right) was performed. Bars show mean ± SEM of three independent experiments in duplicates. *P-value ≤ 0.05, Student’s t-Test. The scale bar in light microscopy panel 1 is 100 μm. d Cells were transfected with the indicated ASOs at 50 nM. At 48 h after transfection, genomic DNA of these cells was prepared with the Apoptotic DNA Ladder Kit (Roche). The control DNA is from apoptotic U937 cells provided with the kit