Conidia of strain BD1181 were cultured in uncoated glass-bottom dishes (Ibidi® GmbH, Germany; 2.5 mL of medium per well) for 16 h at 25°C (Peñalva, 2005 (link)). After this period, the medium was replaced with fresh medium supplemented with 100 mM L -threonine (Sigma) in place of D -glucose to induce alcA(p)::GFP::fluG expression.
Fluorescence images were acquired using a Zeiss Axio Observer Z1 inverted microscope equipped with a 63x Plan Apochromat 1.4 oil immersion Lens, Axiocam MRm Rev.3 camera, a Zeiss HXP 120C external light source for epifluorescence excitation and fitted with filter set 38HE for green fluorescence (Ex BP 470/40; FT 495; Em BP 525/50) and filter set 43HE for red fluorescence (Ex BP 545/25; FT 570; Em BP 605/70). Numerous samples were observed before taking representative images. Fluorescence levels were measured using Fiji software (Schindelin et al., 2012 (link)).
Fluorescence images were acquired using a Zeiss Axio Observer Z1 inverted microscope equipped with a 63x Plan Apochromat 1.4 oil immersion Lens, Axiocam MRm Rev.3 camera, a Zeiss HXP 120C external light source for epifluorescence excitation and fitted with filter set 38HE for green fluorescence (Ex BP 470/40; FT 495; Em BP 525/50) and filter set 43HE for red fluorescence (Ex BP 545/25; FT 570; Em BP 605/70). Numerous samples were observed before taking representative images. Fluorescence levels were measured using Fiji software (Schindelin et al., 2012 (link)).