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Figure 3 | BMC Genomics

Figure 3

From: Acetylation of p65 at lysine 314 is important for late NF-κB-dependent gene expression

Figure 3

Characterization of specific antibodies against p65 acetylated at K314 or 315. (A) Purified recombinant p65 wild type and the acetylation-deficient mutants were incubated with recombinant p300 in the presence (+) or absence (-) of acetyl Co-A. Proteins were resolved on SDS-PAGE and analyzed by western blot using the indicated antibodies. (B) HEK 293T cells were transfected with p65 wild type or mutants, with (+) or without (-) p300 co-transfection. Acetylation of p65 at specific lysines was assessed by western blot using the specific antibodies. * indicates a non-specific band. (C and D) p65 was immunprecipitated from untreated or TNFα-stimulated cells using the indicated antibodies. Immunoprecipitated proteins were separated by 10% SDS-PAGE and subsequently analyzed by western blot using an anti-p65 antibody. * indicates IgG-band.

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