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Fig. 9 | Cell Communication and Signaling

Fig. 9

From: The intrinsically disordered region of GCE protein adopts a more fixed structure by interacting with the LBD of the nuclear receptor FTZ-F1

Fig. 9

GCEC and LBD FTZ-F1 interact when co-expressed in COS-7 cells. COS-7 cells were transfected with appropriate vectors and pulled-down with ANTI-FLAG M2 Affinity gel. Finally, the samples were analyzed for the presence of GCEC and LBD FTZ-F1 by Western blotting and detected by anti-GFP antibodies. Bands corresponding to YFP-GCEC-FLAG and CFP-LBD FTZ-F1 are marked with arrows. a Transfection with the pECFP-C1/LBD FTZ-F1 expression vector. LBD FTZ-F1 is present in cells lysate (TOT), supernatant (SUP) and in the fraction not bound to gel. No protein was observed in the elution fraction. LBD FTZ-F1 does not bind to the ANTI-FLAG M2 Affinity gel. b Transfection with the pEYFP-C1/GCEC-FLAG expression vector. GCEC is present in cells lysate (TOT), supernatant (SUP) and the elution fraction. GCEC tagged with FLAG binds to the ANTI-FLAG M2 Affinity gel. c Simultaneous transfection with the pECFP-C1/LBD FTZ-F1 and pEYFP-C1/GCEC-FLAG expression vectors. Both proteins (GCEC and FTZ-F1) are detected in the elution fraction. As LBD FTZ-F1 does not possess the FLAG-tag, direct binding to the ANTI-FLAG M2 Affinity gel is not possible and requires interaction with the GCEC-FLAG

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Cell Communication and Signaling

ISSN: 1478-811X