Figure 2

a Western blots with RPS2 antibodies of crude cell extracts from PC-3ML cells following treatment with (top lane) 20 nM DNAZYM-1P for increased times of (lanes 1–6) 0, 8, 12, 24, 32 and 48 hr, respectively. Control cells were treated for identical times with (middle lane) 20 nM scrambled oligonucleotide. (bottom lane) beta actin antibody blots. 2b. Comparisons of the ratio of RPS2/actin from densitometry scans of the Western blots in fig. 2a. 2c. RT-PCR assays showing the relative level of RPS2 expression in (P) PC-3ML; (L) LNCaP; (IR) pBABE-IBC-10a-c-myc; and (C) CPTX-1532 cells (at 90% confluent) which were (□) untreated or treated with (╪) scrambled oligonucleotide, and (░) 2 and (■) 4 ug/ml DNAZYM-1P for 8 hr. Shows that the DNAZYM-1P knocks out RPS2 mRNA expression in all 4 cell lines. (×) NPTX-1532 express low levels of RPS2 mRNA (value set at 1). RT-PCR vales were normalized relative to 18S RNA, and then the fold expression calculated relative to values for untreated NPTX-1532 cells which were set at 1. Results averaged from 3 experiments +/-1 S.D.