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. 2016 Aug 15;27(16):2598–2612. doi: 10.1091/mbc.E16-01-0049

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© 2016 Mellado et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 6: — Analysis of mutations in the protease domain of SltB. (A) Construction of strains expressing the fragment between codons 692 and 1272, which encodes the protease domain of SltB. SM indicates selectable marker. Right, immunodetection of Myc₃- and GFP-tagged versions of SltB^692–1272. Bottom, immunodetection of SltA-HA₃ forms in a strain (MAD3750) expressing SltB^692–1272. (B) Scheme illustrating the SltB forms expressed by the constructed diploids. (C) Immunodetection of SltB forms in haploids and diploids. Coexpression of SltB53-GFP and SltB^692–1272-Myc₃ does not result in proteolysis of SltB53. Coexpression of SltB53-GFP and the wild-type SltB results in low levels of the SltB53^Prot-GFP form (right, longer exposure). In the experiment on the left, a mixture of anti-GFP and anti-Myc primary antibodies was used to visualize both types of SltB-tagged proteins simultaneously.