Figure 3.

Cdc42 is required for proper localization of glucanases Eng1 and Agn1 during cell separation. (a). Sum projections of cells displaying GFP tagged glucanases Eng1 and Agn1 at permissive temperature (25°C) in cdc42⁺ and cdc42-1625 mutants, scale bar-5 µm. (b). Representative images showing localization of Eng1-GFP and Agn1-GFP in cdc42⁺ cells during cell separation, along with respective bright-field images. The pink boxes highlight the 3D-reconstructions of the different localization patterns of Eng1-GFP and Agn1-GFP observed in cdc42⁺ cells: (i) Ring, (ii) Ring + Dot, (iii) Disk. (c). Representative images showing fluorescence images of Eng1-GFP and Eng1-GFP in cdc42-1625 cells along with the respective bright-field images and 3D-reconstructed division site of Eng1-GFP Agn1-GFP localization during cell separation at 25°C. The pink boxes highlight the 3D-reconstructions of the different localization patterns of Eng1-GFP and Agn1-GFP observed in cdc42-1625 cells: Scale bar for all cells is 5 µm; Scale bar for all 3D- reconstructed division site is 2 µm. (d). Quantification of Eng1-GFP and Agn1-GFP localization patterns at the division site in the strains mentioned, n = 3 replicates with ≤28 cells each