Figure 2
Figure 2. Mx-Cre–mediated deletion of Stat5a/b abolishes CML-like MPN induced by BCR-ABL1. (A) Kaplan-Meier survival curve for recipients of p210MIGFP-transduced BM from Mx-Cre;Stat5a/bfl/+ donors (solid line), and for recipients of p210MIGFP-transduced BM from Mx-Cre;Stat5a/bfl/− donors either untreated (dotted line) or treated (dashed line) with pIpC after transplantation as described in the “Methods.” The symbols indicate individual recipient mice, with the disease phenotype of each designated by the shading. No recipients in the Mx-Cre;Stat5a/bfl/− + pIpC cohort developed hematologic disease. (B) PB leukocyte counts at day 50 after transplantation in untreated or pIpC-treated recipients of p210MIGFP-transduced BM from Mx-Cre;Stat5a/bfl/+ (left) or Mx-Cre;Stat5a/bfl/− (right) donors. (C) Flow cytometric plot of GFP expression in PB myeloid cells from 2 representative recipients (mice #39 and #38) of p210MIGFP-transduced BM from Stat5a/bfl/− donors. (D) Southern blot analysis of the extent of recombination of the floxed Stat5a/b allele in genomic DNA from PB leukocytes at day 50 after transplantation. Nomenclature is as in supplemental Figure 1C. The small amount of wild-type Stat5a/b allele in recipients of Stat5a/bfl/− BM (mice #35-40) represents contribution from radioresistant host lymphocytes. (E) Spleen weights at autopsy of untreated or pIpC-treated recipients of p210MIGFP-transduced BM from Mx-Cre;Stat5a/bfl/+ (left) or Mx-Cre;Stat5a/bfl/− (right) donors. (F) Western blot analysis of primary myeloerythroid cell extracts from a representative untreated recipient of p210MIGFP-transduced BM from Mx-Cre;Stat5a/bfl/− donors that developed MPN and from 4 healthy pIpC-treated recipients. As controls, extracts from parental and BCR-ABL1–expressing Ba/F3 cell lines and BM from a nontransplanted Stat5 wild-type mouse were included. Proteins were immunoblotted with the indicated Abs against total or phosphorylated BCR-ABL1, STAT5, and CrkL. An anti-eIF4e immunoblot demonstrating equivalent protein loading is shown at the bottom.

Mx-Cre–mediated deletion of Stat5a/b abolishes CML-like MPN induced by BCR-ABL1. (A) Kaplan-Meier survival curve for recipients of p210MIGFP-transduced BM from Mx-Cre;Stat5a/bfl/+ donors (solid line), and for recipients of p210MIGFP-transduced BM from Mx-Cre;Stat5a/bfl/− donors either untreated (dotted line) or treated (dashed line) with pIpC after transplantation as described in the “Methods.” The symbols indicate individual recipient mice, with the disease phenotype of each designated by the shading. No recipients in the Mx-Cre;Stat5a/bfl/− + pIpC cohort developed hematologic disease. (B) PB leukocyte counts at day 50 after transplantation in untreated or pIpC-treated recipients of p210MIGFP-transduced BM from Mx-Cre;Stat5a/bfl/+ (left) or Mx-Cre;Stat5a/bfl/− (right) donors. (C) Flow cytometric plot of GFP expression in PB myeloid cells from 2 representative recipients (mice #39 and #38) of p210MIGFP-transduced BM from Stat5a/bfl/− donors. (D) Southern blot analysis of the extent of recombination of the floxed Stat5a/b allele in genomic DNA from PB leukocytes at day 50 after transplantation. Nomenclature is as in supplemental Figure 1C. The small amount of wild-type Stat5a/b allele in recipients of Stat5a/bfl/− BM (mice #35-40) represents contribution from radioresistant host lymphocytes. (E) Spleen weights at autopsy of untreated or pIpC-treated recipients of p210MIGFP-transduced BM from Mx-Cre;Stat5a/bfl/+ (left) or Mx-Cre;Stat5a/bfl/− (right) donors. (F) Western blot analysis of primary myeloerythroid cell extracts from a representative untreated recipient of p210MIGFP-transduced BM from Mx-Cre;Stat5a/bfl/− donors that developed MPN and from 4 healthy pIpC-treated recipients. As controls, extracts from parental and BCR-ABL1–expressing Ba/F3 cell lines and BM from a nontransplanted Stat5 wild-type mouse were included. Proteins were immunoblotted with the indicated Abs against total or phosphorylated BCR-ABL1, STAT5, and CrkL. An anti-eIF4e immunoblot demonstrating equivalent protein loading is shown at the bottom.

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