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. Author manuscript; available in PMC: 2025 Feb 22.
Published in final edited form as: FEBS Lett. 2008 Aug 8;582(20):3080–4. doi: 10.1016/j.febslet.2008.07.055

Figure 3. Oct-1 binding to the ORF50 promoter is required for the HMGB1/Rta synergistic response.

Figure 3

(a) Ni-NTA bead bound HMGB1 or beads alone were incubated with 35S-Met-labelled Oct-1, Rta or empty vector control produced by ITT. Following washes, bound proteins were separated by SDS-PAGE and the dried gel was exposed to autoradiograph film for 16 hrs. (b) PCR amplification of the ORF50 promoter from HMGB1-specific or no antibody immunoprecipitates using cell extracts transfected with the pGFP, pHMGB1 and/or pRta-GFP in the presence of pORF50ΔOct. (c) 293T cells were co-transfected with (a) pORF50Δ6 or (b) pORF50ΔOct in the presence of pGFP, pHMGB1 and pRta-GFP as indicated. Cells were harvested 48 hrs post-transfection and cell lysates were assayed for luciferase activity. The variations between 3 replicated assays are indicated and data is presented as fold activation versus empty vector control.