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. 2009 Mar 1;20(5):1280–1288. doi: 10.1091/mbc.E08-10-1005

Figure 3.

Figure 3.

STAT1/STAT3 heterodimers mediate HO-1 promoter activation. Human primary macrophages were incubated for 6 h with AC-CM with or without the addition of 20 μM fludarabine (A) or 10 μM STA-21 (B). MΦ-CM was harvested and added to fresh macrophages for 18 h with or without the addition of fludarabine (A) or STA-21 (B). Western blots are representative for at least three individual experiments. (C) Putative STAT binding sites in the human HO-1 promoter are shown. (D) EMSA analysis using 250 fmol of the oligonucleotides resembling the putative STAT binding site at −2361 to −2369 of the human HO-1 promoter. (E) Competitive EMSA analysis using 2.5 or 25 pM of unlabeled oligonucleotides specific for STAT1, STAT3, or oligonucleotides for the putative STAT binding site at −2361 to −2369, which contained a STAT3 ptm, in addition. One representative EMSA out of three is displayed. (F) HO-1 promoter activity in macrophages after transfection of the corresponding promoter constructs and stimulation with MΦ-CM for 18 h. Histograms show firefly luciferase activity normalized to Renilla luciferase activity. Data represent means ± SEM of at least four independent experiments. Asterisks mark statistically significant differences (p ≤ 0.05).