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. 2007 Sep 19;2(9):e902. doi: 10.1371/journal.pone.0000902

Figure 3. Basonuclin is associated with RPA194 and RPA116, subunits of Pol I.

Figure 3

A, Immunoprecipitations detected a basonuclin-RPA194/RPA116 complex. Immunoprecipitations (IP) were performed with extract of HaCaT cells cultured to a high density and analyzed by Western blots. The IP antibodies are indicated above the gel image and that used for Western blots on the left. B, Basonuclin-deficiency reduces the association of RPA194 and rDNA promoter. Basonuclin was knocked-down in HaCaT cells by siRNA and the cells were analyzed by Western for protein levels and by ChIP for association of various factors to rDNA promoter. The Western (upper panel) showed that basonuclin level was reduced to below detection by treatment of siBnc1-s, but the level of RPA194 and UBF was not affected. The lower panel shows the results of PCR analysis of the amount of rDNA promoter precipitated by ChIP. The siRNA reagents the same as indicated in the upper panel. The Western and ChIP antibodies are indicated on the left. C, A quantification of rDNA promoter precipitated by anti-RPA194 in the presence and absence of basonuclin. Shown are qPCR results normalized to the rDNA quantity in the presence of basonuclin. An asterisk indicates p<0.05 (n = 3).