Figure 3

Tax-2 and NF-YB interact in vivo. Panel A- 293T cells were transiently co-transfected with pcMycNF-YB (2 ug), pcTax-2 V5 (4 ug) and pcfCIITA (3 ug) vectors. Cell extracts were immunoprecipitated (IP) with the anti-V5 monoclonal antibody and the purified complexes were immunoblotted (WB) with the indicated antibodies for the detection of NFYB and CIITA. The expression of the proteins in whole cell extracts was also examined by WB (input) with antibodies directed against myc, Flag and V5. Panel B. 293T cells were transiently transfected with pcTax-2 V5 (4 ug), pcfCIITA (3 ug) and/or the empty vector pcDNA and immunoprecipitated as in A. The purified immunocomplexes were immunoblotted with the anti-NFYB and the anti-Flag antibodies for the detection of the endogenous NFYB and of CIITA, respectively. The expression of the proteins in whole cell extracts was also examined by WB (input) with antibodies directed against NFYB, Flag and V5.