Deoxyribonucleic acid

Plates were prepared with agarose medium consisting of tryptose (Oxoid, Thermo Scientific, Leicestershire, UK), deoxyribonucleic acid (Sigma, St. Louis, MO, USA), sodium chloride (VWR Chemicals, Monroeville, PA, USA) and agar. The pH of the medium was set at 7.3 ± 0.2 and autoclaved at 121 °C for 15 min.
The method was based on a previously published study [24 (link)]. From the S. aureus colonies grown in TSA medium at 37 °C overnight, each of the 18 strains was inoculated with a sterile loop on the surface of DNase agar. The plates were inverted and incubated at 37 °C for 18–24 h. After incubation, 1N of HCL was added to the surfaces of the plates.
S. aureus ATCC 25923 and S. saprophyticus ATCC 15305 were used as positive and negative controls, respectively.

Deoxyribonucleic acid from human placenta (D3035), actin from rabbit muscle (A2522), myosin calcium activated from rabbit muscle (M1636), glyceryl trioleate (T7140), cytidine (C4654) were purchased in Sigma-Aldrich and used without any purification in powder form.

The U. maydis strains used in this work (Table S3) derive from the lab strains AB33 [14 (link)], SG200 [7 (link)], and SG200-pit1∆ [23 (link)]. The cells were grown in complete medium (CM: 0.25% w/v cas-amino acids (Difco), 0.1% w/v yeast extract (Difco), 1.0% v/v vitamin solution, and 6.25% v/v salt solution from Holliday [24 ], 0.05% w/v deoxyribonucleic acid from herring sperm, and 0.15% w/v NH₄NO₃ adjusted to pH 7.0 with NaOH (Sigma Aldrich, Darmstadt, Germany) and supplemented with 1% glucose. The transformation of U. maydis protoplasts followed the protocol from [25 (link)]. The constructs were integrated using homologous recombination into the pep4 or upp3 locus, and the transformants were selected on the corresponding antibiotics (200 μg/mL hygromycin, 150 μg/mL nourseothricin, 2 μg/mL carboxin (Sigma Aldrich, Darmstadt, Germany)).
For fluorescence analysis with plate readers, the strains were grown overnight in CM-glucose, before shifting the cultures to nitrate minimal medium (NM)-glucose for the induction of filamentous growth in AB33. Hyphal growth was induced by changing the media to nitrate minimal medium (NM) supplemented with 1% glucose or arabinose. All strains were confirmed using Southern blot analysis [8 (link)] or genotyping PCR.