Ergosterol

For analyzing intracellular sterols, mycelial samples were heat-dried at 80°C and then ground into fine powder for extraction. Extraction and analysis were performed as previously described (Sun et al., 2013 (link)) using ergosterol (J&K Scientific Ltd.) and KTC standards.
For analyzing extracellular sterols, the cultured medium was extracted by chloroform after lyophilization. The extractions were then analyzed by HPLC-MS as described (Sun et al., 2013 (link)).
The significance of the differences between each two samples were evaluated by an unpaired two-tailed Student’s t-test using GraphPad Prism 7.0 and a p-value of less than 0.05 was considered as significant.

Samples for sterol extraction were prepared as that for RNA extraction described above. Mycelial samples were heat-dried at 80 °C and then ground into fine powder using a mortar and pestle. About 10 mg of mycelial powder was used for sterol extraction in Agilent vials (2 mL). Extraction and chemical analysis of sterols and KTC were performed as previously described [36 (link)] using ergosterol (J&K Scientific, Beijing, China) and KTC standards. The statistical analysis was conducted as previously described [36 (link)].

The samples for sterol extraction were prepared by the same method as for RNA extraction. In order to obtain more stable results of sterol profiles, samples with 24-h KTC treatment were used for the subsequent analysis. The collected mycelia of different strains were heat dried at 80°C and then ground into fine powder for sterol extraction and HPLC-MS analysis as reported previously (29 (link)). Ergosterol (J&K Scientific Ltd., China) and KTC standards were run for comparison.