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. 2017 Mar 1;130(5):903–915. doi: 10.1242/jcs.195263

Fig. 7.

Fig. 7.

Lack of N-myristoylation is not sufficient but binding to glycogen is essential for Stbd1 targeting to MAMs. (A–D) HeLa cells co-transfected with (1-25G2A)–EGFP–Myc and mCherry–Sec61β were immunostained for Myc (A) and TOM20 (C). mCherry red fluorescence corresponding to the ER is shown in B. The (1-25G2A)–EGFP–Myc chimeric protein shows a stronger colocalization with the ER compared to mitochondria (D, thresholded Manders' coefficient, mean±s.e.m.: mitochondria, 0.720±0.018; ER, 0.861±0.015; n=22). (E–G) Representative images of HeLa cells transfected with Stbd1(G2A/W273G)–Myc stained for Myc (E) and glycogen (F); an overlay is shown in G. (H–K) HeLa cells transfected with Stbd1(G2A/W273G)–Myc were stained for Myc (H) and TOM20 (J). ER was labelled using mCherry–Sec61β (I). Stbd1(G2A/W273G) exhibits stronger colocalization with the ER compared to mitochondria (K, thresholded Manders' coefficient, mean±s.e.m.: mitochondria, 0.737±0.025; ER, 0.852±0.012; n=11). For the above, representative images are shown. Asterisks in E–G indicate untransfected cells. ***P<0.0001 (unpaired Student's t-test). Scale bars: 10 µm.