Figure 1
Figure 1. Attenuated formation and self-renewal capacity of granulocyte-macrophage progenitors from GATA-2+/− bone marrow. Bone marrow–nucleated cells from each genotype were plated in colony-forming medium supplemented with myeloid and erythroid growth factors and were assessed for granulocyte (CFC-G), macrophage (CFC-M), granulocyte-macrophage (CFC-GM), erythroid (E), and mixed (CFC-mix) colony formation. Three replicates were used per genotype in each experiment. Individual colony types were tallied on day 10. Representative colony types are depicted (A). May-Grünwald-Giemsa staining was used to confirm the lineage identity of individual colonies; typical morphologies of CFC-M and CFC-GM colonies from each genotype are depicted (B) (magnification ×40). Red arrows show the G lineage, and black arrows depict the M lineage. (A,B) The images were captured by the Axiovert 25 microscope (10× objective lens and a 4× eye piece; Zeiss, Welwyn Garden City, United Kingdom) using Immersol 518N mounting medium (Zeiss), May-Grünwald (Sigma, Dorset, United Kingdom) and Giemsa (Sigma) stains, a QIcam camera (QImaging, Pleasanton, CA), and Open Lab software (version 5.5; Coventry, United Kingdom). The cumulative score of specific lineages is shown (C) (n = 4; P = .01 for CFC-GM; CFC-G, P = .37; CFC-M, P = .08; BFU-E, P = .17; and CFC-mix, P = .43). To test the in vitro self-renewal potential of CFC-GMs from each genotype, marrow from GATA-2+/+ and GATA-2+/− animals was plated in granulocyte-macrophage–specific colony-forming medium. After 10 days, CFC-GM colonies were enumerated and replated into fresh granulocyte-macrophage-specific colony-forming medium. Secondary CFC-GM colonies were scored after 10 days in culture. The replating process was repeated to produce tertiary CFC-GM colonies. Three replicates were used per genotype in each experiment. The CFC-GM sequential replating potential of each genotype is depicted (D) (blue bar indicates GATA-2+/+; purple bar, GATA-2+/−) (1° n = 3, P = .02; 2° n = 3, P = .01; 3° n = 3, P = .04). Three replicates were used per genotype in each experiment. CFC-GM colonies were scored after 10 days in culture. ** indicates statistical significance. Error bars indicate SEM. All statistical analyses were performed using the paired Student t test.

Attenuated formation and self-renewal capacity of granulocyte-macrophage progenitors from GATA-2+/− bone marrow. Bone marrow–nucleated cells from each genotype were plated in colony-forming medium supplemented with myeloid and erythroid growth factors and were assessed for granulocyte (CFC-G), macrophage (CFC-M), granulocyte-macrophage (CFC-GM), erythroid (E), and mixed (CFC-mix) colony formation. Three replicates were used per genotype in each experiment. Individual colony types were tallied on day 10. Representative colony types are depicted (A). May-Grünwald-Giemsa staining was used to confirm the lineage identity of individual colonies; typical morphologies of CFC-M and CFC-GM colonies from each genotype are depicted (B) (magnification ×40). Red arrows show the G lineage, and black arrows depict the M lineage. (A,B) The images were captured by the Axiovert 25 microscope (10× objective lens and a 4× eye piece; Zeiss, Welwyn Garden City, United Kingdom) using Immersol 518N mounting medium (Zeiss), May-Grünwald (Sigma, Dorset, United Kingdom) and Giemsa (Sigma) stains, a QIcam camera (QImaging, Pleasanton, CA), and Open Lab software (version 5.5; Coventry, United Kingdom). The cumulative score of specific lineages is shown (C) (n = 4; P = .01 for CFC-GM; CFC-G, P = .37; CFC-M, P = .08; BFU-E, P = .17; and CFC-mix, P = .43). To test the in vitro self-renewal potential of CFC-GMs from each genotype, marrow from GATA-2+/+ and GATA-2+/− animals was plated in granulocyte-macrophage–specific colony-forming medium. After 10 days, CFC-GM colonies were enumerated and replated into fresh granulocyte-macrophage-specific colony-forming medium. Secondary CFC-GM colonies were scored after 10 days in culture. The replating process was repeated to produce tertiary CFC-GM colonies. Three replicates were used per genotype in each experiment. The CFC-GM sequential replating potential of each genotype is depicted (D) (blue bar indicates GATA-2+/+; purple bar, GATA-2+/−) (1° n = 3, P = .02; 2° n = 3, P = .01; 3° n = 3, P = .04). Three replicates were used per genotype in each experiment. CFC-GM colonies were scored after 10 days in culture. ** indicates statistical significance. Error bars indicate SEM. All statistical analyses were performed using the paired Student t test.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal