![Figure 2. A domain of 50 amino acids in PIAS3 (PIAS382-132) is associated with both MITF and STAT3. (A) Schematic representation of the full-length mouse PIAS3 (PIAS31-628), including the SAP domain, PINIT motif, the zinc finger motifs, and the 2 domains of PIAS3 investigated in this work. (B) Fragments PIAS382-132 and full-length PIAS3 (PIAS31-628) labeled with [35S]methionine in vitro were added to GST-MITF or GST-STAT3 that was immobilized on glutathione-sepharose beads. Retained [35S]-labeled PIAS31-628 or PIAS382-132 was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography. Input and GST alone are shown as control. One representative of 3 experiments is shown. (C) PIAS31-628 labeled with [35S]methionine in vitro was added to GST-MITF or GST-STAT3 that was immobilized on glutathione-sepharose beads. Increasing concentrations of in vitro-translated unlabeled PIAS382-132 or PIAS3443-585 was added to the mixtures. Retained [35S]-labeled PIAS31-628 was determined by SDS-PAGE and autoradiography. One representative of 3 experiments is shown.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/107/7/10.1182_blood-2005-08-3325/5/zh80070693320002.jpeg?Expires=1736047664&Signature=WGQi6Q~j0ZkyIlkWvDHeymZgh1qz~sgYANZQYEVf~mn2uN-NhveKHwe1CSgD5EIxQqumPxgNhLz0addiNAj5VcZgW6ZHldL~8FCSCKVy1WSUtwNzNjNvrW76myxPK3S8fG-MjOgs8KRFhffH6gV~BTPTBbP3obARQSJuGSsvFnI4FsBm8a~nIabsLm-qwPTGjSRPnyxUqJHDbuvtRvsfImvFLGZxf-ONgj1LGet1JXWI~30f4liYmFh4dElwemSw-hkfERsPfVwG4Yxe6f1mDppCKTQL4qe-Z6HCtNihJ99j8H1D~Il2p3U0MlgLkERlB8KsHrYCper-ae55UDzkKA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
A domain of 50 amino acids in PIAS3 (PIAS382-132) is associated with both MITF and STAT3. (A) Schematic representation of the full-length mouse PIAS3 (PIAS31-628), including the SAP domain, PINIT motif, the zinc finger motifs, and the 2 domains of PIAS3 investigated in this work. (B) Fragments PIAS382-132 and full-length PIAS3 (PIAS31-628) labeled with [35S]methionine in vitro were added to GST-MITF or GST-STAT3 that was immobilized on glutathione-sepharose beads. Retained [35S]-labeled PIAS31-628 or PIAS382-132 was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography. Input and GST alone are shown as control. One representative of 3 experiments is shown. (C) PIAS31-628 labeled with [35S]methionine in vitro was added to GST-MITF or GST-STAT3 that was immobilized on glutathione-sepharose beads. Increasing concentrations of in vitro-translated unlabeled PIAS382-132 or PIAS3443-585 was added to the mixtures. Retained [35S]-labeled PIAS31-628 was determined by SDS-PAGE and autoradiography. One representative of 3 experiments is shown.
