Figure 7.
HbF differentially affects HbSC RBC parameters, improving HbSC RBC hydration at low levels and ameliorating sickling at modest levels. (A) Experimental design for panels B through F. HbSC female mice were time-mated with HbSS male mice to get a ∼50:50 ratio of HbSS and HbSC pups. Pregnant females were treated with 25 mg/kg BW HU by oral gavage from day 15 through day 21 of gestation. All pups born from the HU-treated pregnant females were treated with HU 50 mg/kg BW intraperitoneally (IP), 3 days per week for 8 weeks (HU-50 group). Control pregnant females and pups received the same volume of PBS by oral gavage or IP injections, respectively (HU-0 group). A limited amount of blood was collected pragmatically to allow serial analysis on the same set of mice. Hence, microbleeds were done at the indicated time points for quantifying F cells by flow cytometry. When the pups were 4 and 8 weeks old, blood was collected for both F-cell and CBC analyses. The data from these experiments are shown in panels B through F. Experimental design for panels G through I: in 1 experiment, we euthanized pups at P7 to P8 to obtain sufficient blood for HPLC, F cell, and oxygen-gradient ektacytometry. (B) Percent F cells determined using flow cytometry are shown at different ages indicated on the X-axis. (C-D) CBC analysis performed at 4 and 8 weeks is shown. The MCHC (C) and MCV (D)are shown. Percent F cells in 7-to-8-day-old HbSC pups (E) and HbSS (F) treated with the HU-0 and HU-50 groups, determined by flow cytometry, are shown. The HbF, HbC, and HbS percentage by HPLC analysis on the P7 to P8 pups in HbSC (G) and HbSS (H) are shown. Oxygen-gradient ektacytometry performed on the P7 to P8 pups HbSC (I) and HbSS (J) mice treated with HU-0 or HU-50. The PoS is shown on the curve. Each symbol represents an individual mouse, and the bars represent mean ± standard error of the mean. The oxygen-gradient ektacytometry curves represent the means of 3 mice per group. Statistics were performed using ANOVA. ∗∗P < .01; ∗∗∗P < .0001; ∗∗∗∗P < .00001. CBC, complete blood count; d, day; Gp, group; w or wk, week.

HbF differentially affects HbSC RBC parameters, improving HbSC RBC hydration at low levels and ameliorating sickling at modest levels. (A) Experimental design for panels B through F. HbSC female mice were time-mated with HbSS male mice to get a ∼50:50 ratio of HbSS and HbSC pups. Pregnant females were treated with 25 mg/kg BW HU by oral gavage from day 15 through day 21 of gestation. All pups born from the HU-treated pregnant females were treated with HU 50 mg/kg BW intraperitoneally (IP), 3 days per week for 8 weeks (HU-50 group). Control pregnant females and pups received the same volume of PBS by oral gavage or IP injections, respectively (HU-0 group). A limited amount of blood was collected pragmatically to allow serial analysis on the same set of mice. Hence, microbleeds were done at the indicated time points for quantifying F cells by flow cytometry. When the pups were 4 and 8 weeks old, blood was collected for both F-cell and CBC analyses. The data from these experiments are shown in panels B through F. Experimental design for panels G through I: in 1 experiment, we euthanized pups at P7 to P8 to obtain sufficient blood for HPLC, F cell, and oxygen-gradient ektacytometry. (B) Percent F cells determined using flow cytometry are shown at different ages indicated on the X-axis. (C-D) CBC analysis performed at 4 and 8 weeks is shown. The MCHC (C) and MCV (D)are shown. Percent F cells in 7-to-8-day-old HbSC pups (E) and HbSS (F) treated with the HU-0 and HU-50 groups, determined by flow cytometry, are shown. The HbF, HbC, and HbS percentage by HPLC analysis on the P7 to P8 pups in HbSC (G) and HbSS (H) are shown. Oxygen-gradient ektacytometry performed on the P7 to P8 pups HbSC (I) and HbSS (J) mice treated with HU-0 or HU-50. The PoS is shown on the curve. Each symbol represents an individual mouse, and the bars represent mean ± standard error of the mean. The oxygen-gradient ektacytometry curves represent the means of 3 mice per group. Statistics were performed using ANOVA. ∗∗P < .01; ∗∗∗P < .0001; ∗∗∗∗P < .00001. CBC, complete blood count; d, day; Gp, group; w or wk, week.

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