SPOP-CUL3 complex represses γ-globin independent of BCL11A. (A) Immunoblot analysis with indicated antibodies of differentiated HUDEP-2 cells expressing CUL3 sgRNAs. (B) γ-Globin mRNA was measured by RT-qPCR; data are plotted as percentage of γ-globin over γ-globin + β-globin mRNAs. Data are mean ± SD of 2 biological replicates. (C) Alignment of human and mouse SPOP DNA sequences. Yellow indicates SPOP sgRNA#1 sequence. Two mismatched nucleotides upstream of protospacer adjacent motif (PAM) in mSpop cDNA. (D) γ-Globin mRNA was measured by RT-qPCR; data are plotted as percentage of γ-globin over γ-globin + β-globin mRNAs. Data are mean ± SD of 2 biological replicates. **P < .05; ***P < .01, unpaired Student t tests. n.s., not significant.