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. 2013 Aug 22;9(8):e1003564. doi: 10.1371/journal.ppat.1003564

Figure 1. Truncations of A. fumigatus CnaA revealed important domains required for growth and septal localization.

Figure 1

(A) Scheme of cnaA truncations and domain organization. Constructs were expressed with native cnaA promoter and egfp tag at the C-terminus to visualize localization and complementation in the ΔcnaA mutant. Full length CnaA (1–559 aa), CnaA-T1 (1–347 aa including the catalytic domain), CnaA-T2 (1–400 aa including the CnaB-Binding Helix), CnaA-T3 (1–424 aa including the SPRR linker), and CnaA-T4 (1–458 aa including the Ca2+/Calmodulin-Binding Domain) are shown. (B) CnaA localization after 24 h growth is indicated as cytoplasmic, partial, or septal. Radial growth was assessed by inoculating 1×104 conidia on GMM agar after 5 days at 37°C. (C) Radial growth is depicted as mean diameter after 5 days growth in triplicate. (D) Western detection of CnaA-EGFP fusion proteins using anti-GFP polyclonal antibody and peroxidase labeled anti-rabbit IgG secondary antibody. (E) Calcineurin activity was determined using p-nitrophenyl phosphate as a substrate. Data from two separate experiments, each with 6 technical replicates are presented as mean ± SD of nanomoles of pNPP released/min/mg protein.