FIG. 7.
Menin is required for maintenance of Hox gene expression. (A and B) HeLa cells were subjected to three rounds of transfection with siRNAs specific for transcripts encoding components of MLL/HCF complex (indicated at the tops of gel lanes). RNA and protein samples were prepared 3 days after the initial transfection. Proteins were subjected to Western blot analysis with the antibodies indicated to the right of the panels (anti-MLLC [mmC2.1], anti-ASH2L, anti-WDR5, anti-HCF-1C [H12], anti-HCF-2, antimenin [C19], and antiactin [C4], respectively), which showed specific and efficient knockdown of protein expression. RNA samples were reverse transcribed and used for quantitative real-time PCR analysis for HoxA9 and GAPDH expression determined in triplicate. (B) Relative expression of HoxA9 to GAPDH is shown with error bars indicating standard deviations.