Figure 3
IFNβ is a crucial mediator of BV6/TMZ-induced cell death. (a) A172 cells (left) or T98G cells (right) were transiently transfected with small interfering RNA (siRNA) against IFNβ. IFNβ mRNA levels were analyzed after 120 h by qRT-PCR, normalized to 28S rRNA expression and fold increase in mRNA levels are shown. Mean values+S.D. of three independent experiments performed in duplicate are shown. *P<0.05; **P<0.01. (b) A172 cells (left) or T98G cells (right) were transiently transfected with siRNA against IFNβ. Cells were treated for 120 h with 100 μM TMZ and/or 2 μM BV6 (A172) or 4 μM BV6 (T98G) or dimethyl sulfoxide (DMSO). Apoptosis was determined by fluorescence-activated cell sorting (FACS) analysis of DNA fragmentation of PI-stained nuclei. Mean values+S.D. of three to four independent experiments performed in triplicate are shown; *P<0.05; **P<0.01. (c) A172 cells (left) or T98G cells (right) were transiently transfected with siRNA against IFNAR1. IFNAR1 mRNA levels were analyzed after 120 h by qRT-PCR, normalized to 28S rRNA expression and fold increase in mRNA levels are shown. Mean values+S.D. of three to four independent experiments performed in duplicate are shown. *P<0.05; **P<0.01. (d) A172 cells (left) or T98G cells (right) were transiently transfected with siRNA against IFNAR1. Cells were treated for 120 h with 100 μM TMZ and/or 2 μM BV6 (A172) or 4 μM BV6 (T98G) or DMSO. Apoptosis was determined by FACS analysis of DNA fragmentation of PI-stained nuclei. Mean values+S.D. of three independent experiments performed in triplicate are shown; *P<0.05; **P<0.01. (e) A172 cells (left) or T98G cells (right) were transiently transfected with siRNA against IFNAR1. Cells were treated for 120 h with 100 μM TMZ and/or 1 ng/ml IFNβ or DMSO. Apoptosis was determined by FACS analysis of DNA fragmentation of PI-stained nuclei. Mean values+S.D. of four independent experiments performed in triplicate are shown; *P<0.05; **P<0.01
