Figure 1
Figure 1. Stat5 is required in primary murine hematopoietic cells for TEL-PDGFRB–mediated growth. (A) Cytokine-independent colony formation of 14.5-dpc fetal liver cells expressing either TEL-PDGFRB or c-Myc. Data shown are the mean percentage of colonies formed by Stat5abnull/null and Stat5ab+/null cells relative to Stat5ab+/+ cells for a given experimental condition. Error bars represent standard deviation. TPNeo induced formation of 195 ± 18 Stat5ab+/+, 194 ± 19 Stat5ab+/null, and 3 ± 1 Stat5abnull/null G418-resistant colonies per 105 transduced fetal liver cells. Positive control c-Myc induced formation of 159 ± 8 colonies per 105 Stat5abnull/null fetal liver cells plated and 184 ± 31 colonies in Stat5ab+/+ cells. Cells transduced with negative control MSCV-Neo did not form colonies in the absence of cytokines (data not shown). Transduction efficiency of Stat5ab+/+ and Stat5abnull/null fetal liver cells with MSCV-Myc was 14% and 12% for 2 independent experiments. For TPNeo and MSCV-Neo, transduction efficiency of Stat5ab+/+ cells averaged 25% ± 15% and Stat5abnull/null cells averaged 45% ± 16% for 7 independent transductions of each genotype. (B) TEL-PDGFRB induced cytokine independent colony formation in Stat5a+/+ (469 ± 26 colonies per 105 transduced cells), Stat5a+/− (266 ± 40), and Stat5a−/− (163 ± 16) whole bone marrow cells. Data shown are the mean percentage of colonies formed by Stat5a−/− and Stat5a+/− cells relative to Stat5a+/+ cells for a given experimental condition. Error bars represent standard deviation. Transduction efficiency of Stat5a−/− cells averaged 98% ± 17% and 93% ± 18% for Stat5a+/+ bone marrow cells in 3 independent transductions. (C) TEL-PDGFRB induced cytokine-independent colony formation in Stat5b+/+ (602 ± 39 colonies per 105 transduced cells), Stat5b+/− (135 ± 25), and Stat5b−/− (27 ± 11) whole bone marrow cells. Data shown are the mean percentage of colonies formed by Stat5b−/− and Stat5b+/− cells relative to Stat5b+/+ cells for a given experimental condition. Error bars represent standard deviation. Transduction efficiency of Stat5b−/− cells was 99% and 81% for Stat5b+/+ bone marrow cells. (TPNeo indicates TEL-PDGFRB-PGK Neo; MSCV-Myc, MSCV-c-Myc ires GFP; and ND, not done.)

Stat5 is required in primary murine hematopoietic cells for TEL-PDGFRB–mediated growth. (A) Cytokine-independent colony formation of 14.5-dpc fetal liver cells expressing either TEL-PDGFRB or c-Myc. Data shown are the mean percentage of colonies formed by Stat5abnull/null and Stat5ab+/null cells relative to Stat5ab+/+ cells for a given experimental condition. Error bars represent standard deviation. TPNeo induced formation of 195 ± 18 Stat5ab+/+, 194 ± 19 Stat5ab+/null, and 3 ± 1 Stat5abnull/null G418-resistant colonies per 105 transduced fetal liver cells. Positive control c-Myc induced formation of 159 ± 8 colonies per 105Stat5abnull/null fetal liver cells plated and 184 ± 31 colonies in Stat5ab+/+ cells. Cells transduced with negative control MSCV-Neo did not form colonies in the absence of cytokines (data not shown). Transduction efficiency of Stat5ab+/+ and Stat5abnull/null fetal liver cells with MSCV-Myc was 14% and 12% for 2 independent experiments. For TPNeo and MSCV-Neo, transduction efficiency of Stat5ab+/+ cells averaged 25% ± 15% and Stat5abnull/null cells averaged 45% ± 16% for 7 independent transductions of each genotype. (B) TEL-PDGFRB induced cytokine independent colony formation in Stat5a+/+ (469 ± 26 colonies per 105 transduced cells), Stat5a+/− (266 ± 40), and Stat5a−/− (163 ± 16) whole bone marrow cells. Data shown are the mean percentage of colonies formed by Stat5a−/− and Stat5a+/− cells relative to Stat5a+/+ cells for a given experimental condition. Error bars represent standard deviation. Transduction efficiency of Stat5a−/− cells averaged 98% ± 17% and 93% ± 18% for Stat5a+/+ bone marrow cells in 3 independent transductions. (C) TEL-PDGFRB induced cytokine-independent colony formation in Stat5b+/+ (602 ± 39 colonies per 105 transduced cells), Stat5b+/− (135 ± 25), and Stat5b−/− (27 ± 11) whole bone marrow cells. Data shown are the mean percentage of colonies formed by Stat5b−/− and Stat5b+/− cells relative to Stat5b+/+ cells for a given experimental condition. Error bars represent standard deviation. Transduction efficiency of Stat5b−/− cells was 99% and 81% for Stat5b+/+ bone marrow cells. (TPNeo indicates TEL-PDGFRB-PGK Neo; MSCV-Myc, MSCV-c-Myc ires GFP; and ND, not done.)

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