SPOP depletion increases γ-globin protein and mRNA levels in HUDEP-2 cells. (A) SPOP protein domain structure and position of 3 SPOP sgRNAs within the BTB domain. (B) HbF flow cytometry of differentiated cells transduced with indicated sgRNAs. Data are mean ± standard deviation (SD) (n = 2). (C) Immunoblot analysis with indicated antibodies using whole-cell lysates from cell pools transduced with sgRNAs. (D) γ-Globin mRNA was measured by RT-qPCR; data are plotted as percentage of γ-globin over γ-globin + β-globin gene levels. Data are mean ± SD of at least 2 biological replicates. Diff, differentiation; exp, exposure. ***P < .01 from unpaired Student t tests. BCL11A (XL), full length BCL11A.