Fig. 7.
Fig. 7. Identification of the nucleotides involved in B complex formation. / (A) Scan of the murine enhancer GATA region. The sequence of mGATA*C (previously described) and of 5 mutated probes (mGATA *1 to *5), scanning the murine enhancer GATA region, were compared with that of the wild-type murine (mGATA), rat (rGATA), and human (hGATA) equivalent sequences. For each probe, nucleotides different from the mGATA sequence are boxed. (B) EMSA. The different probes described in panel A were then used in EMSA experiments with nuclear extracts from K562 cells. B and A complexes are indicated by arrows. (C) Functional studies by transfection. The nucleotide abrogating B complex formation as shown in panel B (mGATA*5) was introduced in the −899 murine GPIIb promoter construct. The CAT activity of this mutated promoter (−899 mGATA*5) was compared with that of the wild-type murine and human promoters by transfection of HEL, K562, and LIN-175 cells.

Identification of the nucleotides involved in B complex formation.

(A) Scan of the murine enhancer GATA region. The sequence of mGATA*C (previously described) and of 5 mutated probes (mGATA *1 to *5), scanning the murine enhancer GATA region, were compared with that of the wild-type murine (mGATA), rat (rGATA), and human (hGATA) equivalent sequences. For each probe, nucleotides different from the mGATA sequence are boxed. (B) EMSA. The different probes described in panel A were then used in EMSA experiments with nuclear extracts from K562 cells. B and A complexes are indicated by arrows. (C) Functional studies by transfection. The nucleotide abrogating B complex formation as shown in panel B (mGATA*5) was introduced in the −899 murine GPIIb promoter construct. The CAT activity of this mutated promoter (−899 mGATA*5) was compared with that of the wild-type murine and human promoters by transfection of HEL, K562, and LIN-175 cells.

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