Fig 11.

Degradation of AgtA–GFP after CkiA depletion. Strain VIE051, which carries agtA–gfp driven by its own promoter and ckiA-flag, driven by the thiA promoter was grown for 14 h at 30°C before transfer and/or addition of thiamine. For the eight tracks to the left of the panel, mycelia were grown for 14 h on ammonium (NH4⁺ o/n) and transferred to either GABA or GABA + 10 µM thiamine and grown for the time indicated. For the four tracks to the right of the panel mycelia was pre-grown for 14 h on GABA (GABA o/n) and then shifted to urea or urea + 10 µM thiamine for the time indicated. AgtA and its degradation products were monitored with an anti-GFP antibody, CkiA with an anti-FLAG antibody and total loading by an anti-actin antibody. For details see Experimental procedures.