Figure 3. ZOL has no effect on BSP gene transcription rate in Saos-2 cells.

(A) After 48 h of treatment with ZOL (20 μM), nuclear run-on assay was performed by labelling nascent transcripts in vitro with [α-³²P]UTP, and by hybridizing the radiolabelled RNA to immobilized cDNAs for BSP, GAPDH and vector DNA pUC18. (B) Total RNA isolated from the treated and non-treated cells was analysed by real-time RT–PCR to control ZOL-induced up-regulation of BSP mRNA steady-state level. ZOL-induced changes in BSP mRNA levels in Saos-2 cells are given as fold-induction relative to untreated control cells and normalized for variations in GAPDH mRNA levels. These results are representative of three separate experiments. **P<0.0001 represents significant difference compared with control cell cultures.