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Figure 5 | BMC Molecular Biology

Figure 5

From: Characterization of an Oct1 orthologue in the channel catfish, Ictalurus punctatus: A negative regulator of immunoglobulin gene transcription?

Figure 5

In vitro transcribed Oct transcription factors bind the octamer motif. A. EMSA demonstrating the binding of catfish Oct1 to the first octamer motif within the core enhancer. Lane 1, free probe. Lane 2, TNT reaction without template DNA (control). Lane 3, catfish Oct1 TNT product. Lane 4, catfish Oct1 TNT product + cold competitor. Lane 5, catfish Oct1 TNT product + scrambled competitor. Lane 6, catfish Oct1 TNT product + anti-Oct1 rabbit serum. Lane 7, catfish Oct1 TNT product + NRS (anti-Oct1 rabbit pre-bleed serum). Arrow depicts the Oct1 shift. B. EMSA demonstrating the binding of catfish Oct2β to the first octamer motif within the core enhancer. Lane 1, catfish Oct2β TNT product. Lane 2, catfish Oct2β TNT product + cold competitor. Lane 3, catfish Oct2β TNT product + scrambled competitor. Lane 4, catfish Oct2β TNT product + anti-Oct2β rabbit serum. Lane 5, catfish Oct2β TNT product + NRS (anti-Oct1 rabbit pre-bleed serum. C. Detection of 35S-labeled TNT transcribed and translated Oct1 and Oct2β by SDS-PAGE and phosphor imaging. Solid black bands on the left hand side represent the kaleidoscope protein marker (Bio-Rad, Hercules, CA.), 222kDa, 129kDa, 83kDa, 43kDa (from top to bottom).

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