Figure 4. Barrier-to-autointegration factor interaction with laminopathic forms of prelamin A. HEK293 cells were transfected with FLAG-tagged prelamin A constructs (LA-WT, LA-R527H, LA-R482Q, LA-L647R) in combination with GFP-BAF (GFP-BAF) construct. Cotransfected cells were lysed and subjected to coimmunoprecipitation experiments. Total lysates (whole lysate) and immunoprecipitated proteins, FLAG-IP in panel (A) and GFP-IP in panel (B), were subjected to western blotting analysis. FLAG and GFP immunolabeled bands were observed in whole lysates as well as in immunoprecipitated samples, while no protein staining was observed in coimmunoprecipitation samples obtained in absence of anti-FLAG antibody or anti GFP- antibody (A/G) (A and B). In (B), prelamin A staining was observed in whole lysates and in GFP-IP lanes subjected to specific anti-prelamin A detection (prelamin A) and to anti-lamin A detection performed with Abcam antibody, which detects both mature lamin A (70kDa) and prelamin A (74kDa) (prelamin A, lamin A).