Fig. 3.

Multiple A3s interact with Prefoldin complex members. Co-IP of (A) PFD5 or (B) PFD3 with A3 enzymes. Cell lysates (L) from HEK293T cells were used for FLAG immunoprecipitation of PFD5-or PFD3- FLAG. Co-immunoprecipitating HA-tagged A3s were detected in either the NT or +RNAse immunoprecipitation (IP). Higher contrast PFD5 or PFD3 blots are shown below each blot to demonstrate the presence of PFD5 or PFD3 in each sample. Variable levels are due to PFD polyubiquitination and degradation (76). AP-MS results demonstrated an interaction of A3B, A3D, A3F, but not A3G with PFD3 and PFD5 with varying strengths. Consistent with this, the co-IP shows that in the NT or +RNAse condition, there are interactions between A3B, A3F, and A3D with PFD3 and PFD5, but not A3G. Purification control immunoblots are shown in Supplemental Fig. S9.