Figure 3: Experiment to elucidate the membrane topology of Calfacilitin.
From: Calfacilitin is a calcium channel modulator essential for initiation of neural plate development
(a–d) Examples of results obtained. (a,b) HEK293T cells transfected with Calfacilitin-myc1 and stained without detergent, seen by phase contrast (a) and fluorescence (b). Surface staining is seen. (c,d) cells stained with Calfacilitin-myc6 without detergent seen by phase contrast (c) and fluorescence (d)—no signal is apparent. (e) Summary of results of transfections of COS or HEK293T cells with Calfacilitin tagged with a myc epitope in different positions (as shown in Fig. 2b). —denotes no signal, −/+ denotes a very weak signal and ++ denotes strong staining with anti-myc. The deduced topology is shown in Fig. 2b. Scale bar (for a–d), 30 μm.
