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. 2007 Aug 15;35(16):5474–5486. doi: 10.1093/nar/gkm601

Figure 6.

Figure 6.

Self-association of MBNL1 proteins is mediated by the C-terminal region. (A) Two-hybrid analysis using yeast strains transformed with the following binding domain (BD) and activation domain (AD) plasmids: (i) GAL4 DNA-binding domain (BD) plasmids with either p53 (activation control) or full-length MBNL1(MBNL1); (ii) activation domain plasmids with either T-antigen (activation control), MBNL1 (residues 1–382), MBNL1 1–264 (N-terminal region) or MBNL 239–382 (C-terminal region). Functional interactions between the proteins expressed from the BD and AD plasmids results in growth on the Trp− Leu− His− selection plate. (B) Co-immunopurification of MBNL1 requires the C-terminal region. HEK293T cells were co-transfected for 24 h with plasmids expressing tagged versions of either full-length or N-terminal MBNL1 (V5-MBNL1 FL alone, co-transfected V5-MBNL1 FL and MBNL1 FL-myc, co-transfected V5-MBNL1 FL and MBNL1 N-myc). Cell lysates were prepared and the V5-MBNL1 FL protein immunopurified (αV5) followed by SDS–PAGE (50% of IP sample) and immunodetection of MBNL1 FL-myc or MBNL1 N-myc using mAb 9E10 (top panel, input lanes represent 2.5% of total IP) or V5-MBNL1 FL (bottom panel, only the input lanes are shown).