Fig. 3. Identification of HoxTF as NFY. (A) Protein fractionation experiments determine the approximate apparent molecular weight of HoxTF. A schematic of the method used is shown in the left. The binding potential of renatured F9 EC nuclear extract polypeptide fractions was analysed by EMSA using the MyoD probe. Molecular mass ranges of the fractions used are indicated at the top. (B) A schematic of the HoxTF purification scheme and a silver-stained SDS-PAGE gel of the purified proteins is shown on the left. Lanes A1 and A2 contain proteins eluted from the first DNA-affinity column bearing the MyoDm or the MyoD oligonucleotides, respectively. Lane B contains proteins eluted after passage of the A2 eluate over the MyoD affinity column. The purified proteins with molecular masses of 48, 45, and 36 kDa are indicated by arrows. (C) Confirmation of NFY binding to b4Cwt by EMSA. NFY* indicates the complex supershifted by the addition of anti-NFYA antibody (lane 6).

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