Skip to main content
. 2007 May 14;404(Pt 2):197–206. doi: 10.1042/BJ20061297

Figure 4. Mapping the hTAFII68 (NTD) region that interacts with GAPDH.

Figure 4

(A) Schematic representation of the GST–hTAFII68 (NTD) fusion proteins and their ability to bind to GAPDH. Numbers refer to amino acid residues, and binding ability is indicated by + or −. (B) Strong binding of GAPDH to GST–hTAFII68 (106–159). Recombinant GST–hTAFII68 (NTD) deletion mutants were incubated with HEK-293T cell lysates. Following GST pull-down assays, the bound proteins were eluted with SDS loading buffer and analysed by Western blotting with an anti-GAPDH antibody. The positions of molecular mass markers and of GAPDH are indicated. Three independent experiments were performed, all of which gave similar results. Lane 1, 20% input; lane 2, GST alone; lane 3, GST–hTAFII68 (NTD); lane 4, GST–hTAFII68 (1–72); lane 5, GST–hTAFII68 (29–105); lane 6, GST–hTAFII68 (106–159). IB, immunoblotting; Ab, antibody. (C) Coomassie Blue staining of the GST–hTAFII68 deletions. The amounts of GST-fusion proteins utilized in these assays were fractionated on SDS/PAGE (15% gels) and visualized by Coomassie Blue staining. Lane 1, GST alone; lane 2, GST–hTAFII68 (NTD); lane 3, GST–hTAFII68 (1–72); lane 4, GST–hTAFII68 (29–105); lane 5, GST–hTAFII68 (106–159).