A. nidulans strains were grown in yeast extract and glucose (YG) medium or 1% glucose minimal medium (Nayak et al., 2006 (link)), supplemented with 1 mg/ml uracil, 2.4 mg/ml uridine, 2.5 μg/ml riboflavin, 1 μg/ml para-aminobenzoic acid, and 0.5 μg/ml pyridoxine when required. Glufosinate (Sigma) for bar selection was used at a final concentration of 700 μg/ml (Straubinger et al., 1992 ).
For imaging of germlings, spores were resuspended in 0.5 ml 0.01% Tween-80 solution. The spore suspension was diluted at 1:1000 in liquid minimal medium containing appropriate auxotrophic supplements. The spore and media mix (400 μl) was added to an eight-chambered Nunc Lab-Tek II coverglass (ThermoFisher) and incubated at 30°C for 16–20 h before imaging. For imaging of mature hyphae, spores were inoculated on minimal medium plates containing the appropriate auxotrophic supplements and incubated at 37°C for 12–16 h. Colonies were excised from agar plates and inverted on Lab-Tek plates for imaging. For biochemistry, spores were inoculated in YG medium containing the appropriate auxotrophic supplements for 16–20 h at 37°C either shaking at 200 rpm in an Erlenmeyer flask or incubated in 100-mm Petri dishes.
For imaging of germlings, spores were resuspended in 0.5 ml 0.01% Tween-80 solution. The spore suspension was diluted at 1:1000 in liquid minimal medium containing appropriate auxotrophic supplements. The spore and media mix (400 μl) was added to an eight-chambered Nunc Lab-Tek II coverglass (ThermoFisher) and incubated at 30°C for 16–20 h before imaging. For imaging of mature hyphae, spores were inoculated on minimal medium plates containing the appropriate auxotrophic supplements and incubated at 37°C for 12–16 h. Colonies were excised from agar plates and inverted on Lab-Tek plates for imaging. For biochemistry, spores were inoculated in YG medium containing the appropriate auxotrophic supplements for 16–20 h at 37°C either shaking at 200 rpm in an Erlenmeyer flask or incubated in 100-mm Petri dishes.