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There is an error in panel D of S8 Fig. Specifically, the upper panel should read ‘SclB-cYFP + nYFP’, not ‘SclA-cYFP + nYFP’. The authors have provided a corrected version here.
Supporting information
S8 Fig. GFP-fusion proteins of SclB are functional and phosphorylated and Bi-FC controls are negative.
A) Strains expressing SclB either N- or C-terminally tagged with sGFP in ΔsclB background, ΔsclB and wildtype (WT) were point inoculated on solid MM and grown for 4 days in light. B) SclB-GFP and GFP-SclB fusion proteins expressed under native promoter are visualized in a western hybridization assay employing an α-GFP antibody (GFP) and Ponceau staining as loading control (Pnc). The black arrow indicates bands corresponding to full-length fusion proteins (in silico prediction 87.46 kDa). C) Protein crude extracts of GFP-SclB grown vegetatively were mixed with phosphatase inhibitor cocktail (-/PhoI), with Lambda phosphatase (λ/-), or Lambda phosphatase and phosphatase inhibitor cocktail (λ/PhoI). A control sample was left untreated (-/-). A subsequent western hybridization assay employing α-GFP antibody visualizes protein bands. D) Two strains, either expressing sclB::cyfp and the free second half of the split YFP (nyfp; upper part), or free cyfp and rcoA::nyfp (lower part), under control of a bi-directional nitrate promoter were constructed. Strains were inoculated in liquid MM and analyzed with fluorescence microscopy after 36 h at 30°C.
1.Thieme KG, Gerke J, Sasse C, Valerius O, Thieme S, Karimi R, et al. (2018) Velvet domain protein VosA represses the zinc cluster transcription factor SclB regulatory network for Aspergillus nidulans asexual development, oxidative stress response and secondary metabolism. PLoS Genet
14(7): e1007511
10.1371/journal.pgen.1007511
[DOI] [PMC free article] [PubMed] [Google Scholar]
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Supplementary Materials
S8 Fig. GFP-fusion proteins of SclB are functional and phosphorylated and Bi-FC controls are negative.
A) Strains expressing SclB either N- or C-terminally tagged with sGFP in ΔsclB background, ΔsclB and wildtype (WT) were point inoculated on solid MM and grown for 4 days in light. B) SclB-GFP and GFP-SclB fusion proteins expressed under native promoter are visualized in a western hybridization assay employing an α-GFP antibody (GFP) and Ponceau staining as loading control (Pnc). The black arrow indicates bands corresponding to full-length fusion proteins (in silico prediction 87.46 kDa). C) Protein crude extracts of GFP-SclB grown vegetatively were mixed with phosphatase inhibitor cocktail (-/PhoI), with Lambda phosphatase (λ/-), or Lambda phosphatase and phosphatase inhibitor cocktail (λ/PhoI). A control sample was left untreated (-/-). A subsequent western hybridization assay employing α-GFP antibody visualizes protein bands. D) Two strains, either expressing sclB::cyfp and the free second half of the split YFP (nyfp; upper part), or free cyfp and rcoA::nyfp (lower part), under control of a bi-directional nitrate promoter were constructed. Strains were inoculated in liquid MM and analyzed with fluorescence microscopy after 36 h at 30°C.