Fig. 8. Chitin synthesis and Chs3p distribution after endocytosis blockade. (A) Calcofluor staining and Chs3p-GFP distribution visualized microscopically as described in independent cultures. The indicated strains (BY4742 background) were transformed with the YpLac111-CHS3-GFP plasmid and thus contained two functional copies of Chs3p. Note the uniform distribution of Chs3p-GFP along the plasma membrane in the end4
mutants regardless of the presence of Chs4p. (B) Chs3p-GFP distribution after Latrunculin A treatment. CRM103 (chs3
), CRM892 (chs3
chs4
) and CRM1077 (chs3
chs6
) strains transformed with the YpLac111-CHS3-GFP plasmid were grown to early log phase and treated with 50 µM latrunculin for one hour. (C) Same as in B but with treatment with 15 µM latrunculin for 15 minutes. Note that the fluorescence intensity is not uniform along the cell surface.