![Figure 2. FLT3 phosphorylation in FLT3 TKI-sensitive and -resistant cell lines and primary samples decreases with FLT3 TKI treatment, although STAT5 and/or Akt and/or MAPK pathways remain activated in most resistant cells. Cell lines expressing (A) wt FLT3 [SEM-K2], (B) FLT3-ITD [MOLM14], or (C) FLT3(D835H) [Hb1119] were treated with PBS, 50 nM CEP-5214, 50 nM CEP-701, 50 nM PKC412, or 1 μM AG1295 for 1 hour at 37°C. Arrow points to P-Akt. (D) Indicated cell lines were treated with PBS or 50 nM CEP-701 for 1 hour at 37°C. (E) Primary samples (20 × 106 cells) were treated with PBS, 50 nM CEP-701, or 50 nM PKC412 for 1 hour at 37°C. Immunoprecipitates and total protein extracts were resolved by 8% SDS–polyacrylamide gel electrophoresis (PAGE) or 10% SDS-PAGE, respectively, and subjected to immunoblot analysis with the indicated phospho-specific antibodies. The same blots were then stripped and reprobed with protein-specific antibodies.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/109/4/10.1182_blood-2006-05-023804/4/zh80040708460002.jpeg?Expires=1753346238&Signature=NA9iqw7~J4tWD4mAsQKiMUUpcAhFr4GRBuZrPtbS4J84wF1f7~6PWJW9jzqITaHIh-jo-PSjyBlbbZD~TY~C9T1rau3Rc5nveQp9eqYtEUBsYDvifVz12UvYKm3JWNDLjX1qdH3F1k~8ONXyhtOv1OW0mY0R0PzmSc~m6i52Qs3P7SvmWIz9judQXDNxo~i4G~25dci4MIlFmEWNE~tFamFZ30HMWQNHu9oyeNGmZxEILOOc1k~LMGK32ulTsHnTFnP-hcMkxOcI51~QTyn5D6JqWw7TTmbQGhjm0tQOcnqqaVi873MawZPmF3uHLuF~2CaKQfD1qCIikj6twasLbw__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
FLT3 phosphorylation in FLT3 TKI-sensitive and -resistant cell lines and primary samples decreases with FLT3 TKI treatment, although STAT5 and/or Akt and/or MAPK pathways remain activated in most resistant cells. Cell lines expressing (A) wt FLT3 [SEM-K2], (B) FLT3-ITD [MOLM14], or (C) FLT3(D835H) [Hb1119] were treated with PBS, 50 nM CEP-5214, 50 nM CEP-701, 50 nM PKC412, or 1 μM AG1295 for 1 hour at 37°C. Arrow points to P-Akt. (D) Indicated cell lines were treated with PBS or 50 nM CEP-701 for 1 hour at 37°C. (E) Primary samples (20 × 106 cells) were treated with PBS, 50 nM CEP-701, or 50 nM PKC412 for 1 hour at 37°C. Immunoprecipitates and total protein extracts were resolved by 8% SDS–polyacrylamide gel electrophoresis (PAGE) or 10% SDS-PAGE, respectively, and subjected to immunoblot analysis with the indicated phospho-specific antibodies. The same blots were then stripped and reprobed with protein-specific antibodies.
