FIGURE 7.

Effects of siRNA and shRNA treatment on PCMT1 transcript levels. Panel A, set up of apoptosis conditions. HepG2 were treated with cisplatinum at different times. In the subsequent experiments we chose 35 mm cisplatinum for 6 h (defined as condition A in the text). Panel B, PCMT1 silencing estimated by real time PCR in HepG2 transfected with siRNA and then subject to condition A. C siRNA rd, negative control transfected with scrambled siRNA; siRNA MAPK, positive control (see “Experimental Procedures”); siRNA pcmt3, cell sample transfected with siRNA against PCMT1 3′ UTR; siRNA pcmt5, cell samples transfected with siRNA against PCMT1 5′ UTR. *, p < 0.01. Panel C, PCMT1 silencing estimated by real time PCR in HepG2 transfected with shRNA and then subject to condition A. C shRNA RD, negative control transfected with scrambled shRNA; shRNAvs5′UTRpcmt(a), shRNA directed against the 5′ UTR of PCMT1 transcript; shRNAvs5′UTRpcmt(b), shRNA directed against a different 5′ UTR of the PCMT1 transcript; shRNAvsORFpcmt(a), shRNA directed against the PCMT1 transcript ORF; shRNAvsORFpcmt(b), shRNA directed against a different sequence in the PCMT1 ORF. **, p < 0.01