Fig. 2
Transcriptional upregulation of S100A4 by HIF-1α. A Western blot (left) and RT-PCR analyses (right) for the indicated molecules in total lysates or total RNA from KS-1 cells treated with 100 and 200 μM CoCl2 for 24 h. B The S100A4 promoter sequence containing five putative HIF-1α -binding sites (BEs). C KS-1 cells were transfected with Wild type (WT) 1 (− 1986/+ 1012) and WT 2 (− 447/+ 1012) S100A4-luc reporter constructs, together with HIF-1α. Relative activity was determined based on arbitrary luciferase light units normalized to pRL-TK activity. The activities of the reporter plus the effector relative to that of the reporter plus empty vector are shown as means ± SDs. The experiment was performed in duplicate. D Various promoter constructs with mutations (Mut, mutant-type) in the putative HIF-1α-BEs were used for evaluating transcriptional regulation of the S100A4 promoter by HIF-1α. E KS-1 cells were transfected with various mutant constructs of S100A4 promoter, along with HIF-1α