Figure 5
Figure 5. Early and late stages in erythropoiesis are rescued in Gfi-1−/− mice by reducing Id2 levels. (A) Representative flow cytometric analyses of erythroid development in BMCs using CD71 and Ter119 surface markers. Upper left quadrant of contour plot contains CD71+/Ter119−/lo erythroid-committed progenitors, which further develop into CD71+/Ter119+ erythroblasts in the upper right quadrant of the contour plot, which differentiate into more mature CD71−/Ter119+ erythroid contains in the lower right quadrant of the contour plot. (B) Bar graph showing the percentages of erythroid populations identified by CD71 and Ter119 expression on Gfi-1+/+, Gfi-1−/−, and Gfi-1−/−;Id2+/− BMCs. The percentages can be directly compared between mice, because the total BM cellularity of all mice examined was equivalent. The data are presented as mean ± SD and are representative of 3 independent experiments. *P < .05 for Gfi-1−/− compared with Gfi-1−/−;Id2+/− CD71+/Ter119−/lo and CD71−/Ter119+ cell populations. (C-D) Bar graphs showing the percentages of erythroid populations identified by CD71 and Ter119 expression on (C) Gfi-1+/+;Id1+/−, Gfi-1−/−;Id1+/+, and Gfi-1−/−;Id1+/− BMCs and (D) Gfi-1+/+;Id3+/−, Gfi-1−/−;Id3+/+, and Gfi-1−/−;Id3+/− BMCs. The data are presented as the mean ± SD. (E) Analysis of CD71 and Ter119 expression in c-Kit+ BMCs in Gfi-1+/+, Gfi-1−/−, and Gfi-1−/− ; Id2+/− mice. These include the percentages of CD71med/loTer119−/lo (A), CD71hiTer119med (B), and CD71loTer119lo (C) c-Kit+ BMCs. CD71hiTer119med erythroid progenitors in c-Kit+ BMCs were restored by reducing Id2 levels in Gfi-1−/− mice, and the increase in c-Kit+ CD71loTer119lo cell population in Gfi-1−/− BMCs was reduced by decreasing Id2 levels in vivo. (F) Total number of c-Kit+ CD71med/loTer119−/lo, c-Kit+ CD71hiTer119med, and c-Kit+ CD71loTer119lo erythroid progenitor cells in BMCs using total BM cellularity, percentage of c-Kit+ BMCs, and percentage of erythroid progenitors from 2 femurs and 2 tibias of each mouse. Data are presented as the mean ± SD.

Early and late stages in erythropoiesis are rescued in Gfi-1−/− mice by reducing Id2 levels. (A) Representative flow cytometric analyses of erythroid development in BMCs using CD71 and Ter119 surface markers. Upper left quadrant of contour plot contains CD71+/Ter119−/lo erythroid-committed progenitors, which further develop into CD71+/Ter119+ erythroblasts in the upper right quadrant of the contour plot, which differentiate into more mature CD71/Ter119+ erythroid contains in the lower right quadrant of the contour plot. (B) Bar graph showing the percentages of erythroid populations identified by CD71 and Ter119 expression on Gfi-1+/+, Gfi-1−/−, and Gfi-1−/−;Id2+/− BMCs. The percentages can be directly compared between mice, because the total BM cellularity of all mice examined was equivalent. The data are presented as mean ± SD and are representative of 3 independent experiments. *P < .05 for Gfi-1−/− compared with Gfi-1−/−;Id2+/− CD71+/Ter119−/lo and CD71/Ter119+ cell populations. (C-D) Bar graphs showing the percentages of erythroid populations identified by CD71 and Ter119 expression on (C) Gfi-1+/+;Id1+/, Gfi-1−/−;Id1+/+, and Gfi-1−/−;Id1+/ BMCs and (D) Gfi-1+/+;Id3+/, Gfi-1−/−;Id3+/+, and Gfi-1−/−;Id3+/ BMCs. The data are presented as the mean ± SD. (E) Analysis of CD71 and Ter119 expression in c-Kit+ BMCs in Gfi-1+/+, Gfi-1−/−, and Gfi-1−/− ; Id2+/− mice. These include the percentages of CD71med/loTer119−/lo (A), CD71hiTer119med (B), and CD71loTer119lo (C) c-Kit+ BMCs. CD71hiTer119med erythroid progenitors in c-Kit+ BMCs were restored by reducing Id2 levels in Gfi-1−/− mice, and the increase in c-Kit+ CD71loTer119lo cell population in Gfi-1−/− BMCs was reduced by decreasing Id2 levels in vivo. (F) Total number of c-Kit+ CD71med/loTer119−/lo, c-Kit+ CD71hiTer119med, and c-Kit+ CD71loTer119lo erythroid progenitor cells in BMCs using total BM cellularity, percentage of c-Kit+ BMCs, and percentage of erythroid progenitors from 2 femurs and 2 tibias of each mouse. Data are presented as the mean ± SD.

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