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. 2023 Jun 7;12(12):1580. doi: 10.3390/cells12121580

Figure 5.

Figure 5

Transient transfection of C-terminal domain fragments from BFSP1 confirms 434–460 as a bone fide LBD. A series of eGFP-tagged BFSP1 constructs derived from the C-terminal domain designed to confirm the membrane binding of the amphipathic helix associated with the region 434–452 were subcloned into the peGFP-N3 vector (A). Transient transfection of these different constructs into MCF7 cells produced prominent plasma membrane staining for some (B; 434–665; G434–P548; A434–P548), but not all constructs (B; 461–548, 550–548). Furthermore, preventing myristoylation by replacing the N-terminal glycine residue with alanine (G434A) did not prevent the plasma membrane localization of BFSP1 434–548, suggesting that myristoylation was not necessary for its plasma membrane, or internal membrane localisation. Scale bar = 10 µm (B).