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. 1997 May 13;94(10):5355–5360. doi: 10.1073/pnas.94.10.5355

Figure 2.

Figure 2

Induction of HES-1 in SCLC cells. (A) DMS53 cells containing a tetracycline-inducible transcriptional activator were stably transfected with HES-1 plasmids driven by a tetracycline response element, treated with carrier or doxycycline (1 μg/ml) for various time periods, and analyzed for HES-1 protein expression by Western blotting as described in Fig. 1. Lanes: 1, carrier-treated DMS53-HES-1 (sense orientation HES-1 construct) cells, 48 hr; 2, doxycycline-induced DMS53-HES-1 cells, 48 hr; 3, carrier-treated DMS53-HES-1 cells, 146 hr; 4, doxycycline-treated DMS53-HES-1 cells, 146 hr; 5, carrier-treated DMS53-vector cells, 48 hr; 6, doxycycline-treated DMS53-vector cells, 48 hr; 7, carrier-treated DMS53-rev-HES-1 (antisense orientation HES-1 construct) cells, 48 hr; 8, doxycycline-treated DMS53-rev-HES-1 cells, 48 hr. (B) Effect of HES-1 induction on hASH1 mRNA levels. Total RNA (20 μg) from DMS53 cells transfected with tetracycline-inducible constructs, and treated with carrier (TET−) or 1 μg/ml doxycycline (TET+) for 8, 48, and 146 hr, was analyzed for hASH1 and GAPDH mRNA as described in Materials and Methods. In the bottom panels, the ratios for the relative intensities of the hASH1 and GAPDH hybridization signals were calculated from PhosphorImager values.