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. 2015 Nov 26;25(3):472–483. doi: 10.1093/hmg/ddv486

Figure 5.

Figure 5.

Co-immunoprecipitation of WT and P76L POU1F1 proteins with three cofactors. HEK293T cells were co-transfected with plasmids pcDNA4-POU1F1-HA expressing POU1F1_WT or P76L (lanes labeled WT and P76L, respectively) and with each of the following three expression vectors: pcDNA3-PITX1 (A), pcDNA3-LHX3a-Flag (B) and pcDNA3-ELK1-Flag (C). Nuclear extracts were co-immunoprecipitated with an anti-HA antibody, and the western blots were revealed with an anti-PITX1 antibody for PITX1 and an anti-Flag antibody for LHX3 and ELK1 (left panels). The same nuclear extracts were co-immunoprecipitated with an anti-PITX1 antibody for PITX1 and an anti-Flag antibody for LHX3 and ELK1, and westerns blots revealed with the anti-HA antibody (right panels); IP: immunoprecipitation, WB: western blots. Quantification of the complex formed, represented by histograms, was calculated as the ratio of protein immunoprecipitated to protein expressed (input).