BTN1 overexpression or deletion has opposing effects on Golgi morphology. (A) Deletion of BTN1 enhances Golgi clustering in cells expressing GFP-Sed5. WT, btn1Δ, and WT cells overexpressing BTN1 from a multicopy plasmid (BTN1) and all expressing GFP-Sed5 from a single-copy plasmid are shown. Note the large puncta in btn1Δ cells. (B) Deletion of BTN1 enhances Golgi clustering in cells expressing GFP-Sed5317A, whereas BTN1 overexpression enhances fragmentation (same as in A, except all strains express GFP-Sed5317A). Note the large puncta in WT and btn1Δ cells. (C) Golgi clustering is reduced in btn1Δ cells expressing GFP-Sed5317D (same as in A, except all strains express GFP-Sed5317D). (D) Deletion of BTN1 enhances Golgi clustering in cells expressing GFP-Sed5 from its genomic locus (GFP-Sed5 integrated [int.]). WT, btn1Δ, and WT cells overexpressing BTN1 from a multicopy plasmid (BTN1) and all expressing GFP-SED5 from its genomic locus are shown. Note the larger puncta in btn1Δ cells. (E) BTN1 overexpression reduces Golgi clustering in cells expressing GFP-Sed5317A from its genomic locus (same as in D, except all cells express GFP-SED5317A). Note the reduction in puncta size in cells overproducing Btn1. (F) Deletion of BTN1 enhances Golgi clustering in cells expressing GFP-Sed5317D from its genomic locus (same as in D, except all cells express GFP-SED5317D). Note the enhancement in puncta size in cells lacking BTN1. Bars, 1 µm.
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