Fig. 2.
Fig. 2. IL-6–independent cells express constitutive STAT3 binding to a STAT3/APRF element. / Nuclear extracts were prepared from IL-6–dependent cells cultured in the absence of IL-6 for 10 hours (T1165) or 48 hours (7TD1 and B9) and restimulated with medium alone or IL-6 (100 U/mL) for 20 minutes. IL-6–independent cells were cultured continuously in medium without IL-6 or stimulated with IL-6 (100 U/mL) for 20 minutes. EMSA was performed with the extracts, and a probe was derived from the overlapping APRF/NF-κB site in the junB enhancer region.22 The probe, GGG CAG ATT CCGGGA ATC GAG TCC CCC C, contains an APRF element (bold) that binds STAT3 and a mutation in the overlapping NF-κB site (underlined) to avoid interference with binding to the STAT3 site. The arrow in the second panel points to STAT3-specific band.

IL-6–independent cells express constitutive STAT3 binding to a STAT3/APRF element.

Nuclear extracts were prepared from IL-6–dependent cells cultured in the absence of IL-6 for 10 hours (T1165) or 48 hours (7TD1 and B9) and restimulated with medium alone or IL-6 (100 U/mL) for 20 minutes. IL-6–independent cells were cultured continuously in medium without IL-6 or stimulated with IL-6 (100 U/mL) for 20 minutes. EMSA was performed with the extracts, and a probe was derived from the overlapping APRF/NF-κB site in the junB enhancer region.22 The probe, GGG CAG ATT CCGGGAATCGAG TCC CCC C, contains an APRF element (bold) that binds STAT3 and a mutation in the overlapping NF-κB site (underlined) to avoid interference with binding to the STAT3 site. The arrow in the second panel points to STAT3-specific band.

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