Figure 5.
PUS1 deficiency impairs erythroid development in mice. (A) Complete blood count analysis of WT and Pus1S172fs/S172fs mice (S172fs) aged 4 weeks with different gender. Green dotted lines define the normal ranges. Female mice at 4 weeks: WT, n = 7; S172fs, n = 11. Male mice: WT, n = 8; S172fs, n = 5. (B-D) Flow cytometry analysis of erythroblasts in BM of mice aged 4 weeks. The gating strategy of erythroblasts by flow cytometry. R I to R IV represent proerythroblasts (region I, CD71highTer119int), basophilic erythroblasts (region II, CD71highTer119+), late basophilic and chromatophilic erythroblasts (region III, CD71intTer119+), and orthochromatophilic erythroblasts (region IV, CD71−Ter119+), respectively. Representative graph (B), frequency statistics (C), and absolute numbers (D) of different stages are shown. WT (n = 11), 7 female mice and 4 male mice at 4 weeks; S172fs (n = 9), 7 female mice and 2 male mice at 4 weeks. (E-G) Flow cytometry analysis of erythroblasts in spleen (SP) cells of mice aged 4 weeks. Representative graph (E), frequency statistics (F), and absolute numbers (G) of different stages are shown. WT (n = 11), 7 female mice and 4 male mice at 4 weeks; S172fs (n = 9), 7 female mice and 2 male mice at 4 weeks. (H) Schematic diagram of serial competitive transplant assay. (I) Frequency of donor cells of mature erythrocytes in serial competitive transplant assay. Primary competitive transplantation (n = 7), 4 female mice and 3 male mice; secondary competitive transplantation (n = 6), 3 female mice and 3 male mice. Values in all panels denote mean ± SD (∗P < .05; ∗∗P < .01; ∗∗∗P < .001); unpaired Student t test (A), paired Student t test (I), or 2-way ANOVA (C-D, F-G).

PUS1 deficiency impairs erythroid development in mice. (A) Complete blood count analysis of WT and Pus1S172fs/S172fs mice (S172fs) aged 4 weeks with different gender. Green dotted lines define the normal ranges. Female mice at 4 weeks: WT, n = 7; S172fs, n = 11. Male mice: WT, n = 8; S172fs, n = 5. (B-D) Flow cytometry analysis of erythroblasts in BM of mice aged 4 weeks. The gating strategy of erythroblasts by flow cytometry. R I to R IV represent proerythroblasts (region I, CD71highTer119int), basophilic erythroblasts (region II, CD71highTer119+), late basophilic and chromatophilic erythroblasts (region III, CD71intTer119+), and orthochromatophilic erythroblasts (region IV, CD71−Ter119+), respectively. Representative graph (B), frequency statistics (C), and absolute numbers (D) of different stages are shown. WT (n = 11), 7 female mice and 4 male mice at 4 weeks; S172fs (n = 9), 7 female mice and 2 male mice at 4 weeks. (E-G) Flow cytometry analysis of erythroblasts in spleen (SP) cells of mice aged 4 weeks. Representative graph (E), frequency statistics (F), and absolute numbers (G) of different stages are shown. WT (n = 11), 7 female mice and 4 male mice at 4 weeks; S172fs (n = 9), 7 female mice and 2 male mice at 4 weeks. (H) Schematic diagram of serial competitive transplant assay. (I) Frequency of donor cells of mature erythrocytes in serial competitive transplant assay. Primary competitive transplantation (n = 7), 4 female mice and 3 male mice; secondary competitive transplantation (n = 6), 3 female mice and 3 male mice. Values in all panels denote mean ± SD (∗P < .05; ∗∗P < .01; ∗∗∗P < .001); unpaired Student t test (A), paired Student t test (I), or 2-way ANOVA (C-D, F-G).

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