FIG. 5.
USF activates β-globin gene expression in MEL cells. Stable MEL cell lines were created containing either A-USF (pTO/A-USF) or USF1 (pTO/USF1) and the Tet repressor (pTR). The expression of A-USF and USF1 was induced by incubating the cells with doxycycline and analyzed by Western-blotting experiments (A and C). βmaj-Globin gene expression in transfected cell lines was analyzed by quantitative RT-PCR, using the expression of GAPDH as an internal reference, and compared to the expression in cells transfected with the empty vector (pTO), whose expression level was set to 1. (A and B) Western blot analysis of A-USF expression (A) and analysis of β-globin gene expression (B) in MEL cells transfected with pTO/A-USF or pTO in the presence or absence of doxycycline as indicated. A-USF was detected using an antibody against the HA tag (α-HA), and USF1 was detected with a USF1-specific antibody. Error bars indicate standard deviations. (C and D) Western blot analysis of USF1 expression (C) and analysis of βmaj-globin gene expression (D) in MEL cells transfected with pTO/USF1 or pTO. USF1 was detected with a USF1-specific antibody. Data marked with an asterisk had P values above 0.05. + and +/− are as defined for Fig. 2.