Figure 7.

Expression of TBLR1 alters the expression of SMRT and N-CoR. a. Transient expression of TBLR1 alters the expression of co-transfected SMRT(1-300). COS7 cells were co-transfected with pCDNA His-SMRT(1-300) and the His-tagged TBLR1 constructs described above. Control cells were transfected with pCDNA His-SMRT(1-300) and an empty vector.36 hrs after transfection the cells were lysed and equal amount of proteins were electrophoresed in SDS-PAGE gel. After transfer anti-His mAb was used to detect the transiently expressed proteins. The TBLR1 constructs used in this experiment were also His-tagged and are visible on the blot after staining. They are marked with * in the figure. The immuno-precipitates were quantified by phosphorimaging. After stripping, the blots were restained with anti ERK as a control for loading differences. b. Stable expression of TBLR1 alters the expression of endogenous N-CoR. Jurkat were infected with retroviruses expressing TBLR1 or the TBLR1 [δN] and TBLR1 [δC] construct. The retroviral vector also expressed GFP which was used to identify the infected cells. After two rounds of sorting the cells were analyzed by Western blotting for their expression of N-CoR. Th blot was then stripped and restained with an antibody to α tubulin as a loading control. c. Stable expression of TBLR1 does not alter the expression of endogenous N-CoR or SMRT message. mRNA was prepared from the cell lines used in Figure 3b and examined using quantitative real-time RT-PCR. Primer pairs that identified GAPDH and N-CoR were used. The results shown are representative of 3 experiments performed in triplicate. The samples are labeled as follows: black = control Jurkat; red = Jurkat (TBLR1); blue = Jurkat (TBLR1 [δC]); green = Jurkat(TBLR1 [δN])

Zhang et al. BMC Cell Biology 2006 7:31   doi:10.1186/1471-2121-7-31
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