## 2 Materials and Methods
### Quantification of Nucleic Acid, Protein, Chitin, \(\beta\)-1,3-Glucan, and Ergosterol  
Briefly, 1 mL of the supernatant obtained by centrifugation was tested for nucleic acid and protein leakage after an appropriate dilution by measuring the absorbance at 260 nm and 280 nm. \(\beta\)-1,3-glucan and chitin quantification was performed as described by [PERSON] et al. (2019) [17] and [PERSON] et al. (2018) [18] with some modifications. The mycelium sample (40 mg) was suspended in 2 mL of 1 M NaOH solution, mixed, and incubated at 52 \({}^{\circ}\)C for 30 min. A total of 0.5 mL supernatant was collected in a new tube, and 1.85 mL of an aniline blue mix was added for incubation at 52 \({}^{\circ}\)C for 30 min. Fluorescence was detected using a fluorescence spectrophotometer (HITACHI F-7000) with 405 nm excitation and 460 nm emission. \(\beta\)-1,3-glucan content was expressed as the relative fluorescence unit percentage per milligram of mycelium. To sample 30 mg mycelium, 3 mL of a saturated KOH solution was added and heated at 130 \({}^{\circ}\)C for 1 h. The sample was cooled to room temperature, thereafter, 8 mL chilled 75% (\(v/v\)) ethanol was added, and the sample was shaken well and then incubated in an ice bath for 5 min. Next, 0.9 mL of 13.3% (\(w/v\)) celite545 was added, then the mixture was shaken for 5 min and then centrifuged at 4 \({}^{\circ}\)C at 5000 rpm for 5 min. The precipitate was washed with 10 mL of chilled 40% (\(v/v\)) ethanol and 10 mL of chilled distilled water. The sample was prepared by centrifugation to remove the solution; 20 mL distilled water was then added, and 0.5 mL of the mixture was taken as the sample. A standard solution (10 mg/mL glucosamine) and a blank control (distilled water) were prepared. 0.5 mL of water, 5% (\(w/v\)) NaNO\({}_{2}\), and 5% (\(w/v\)) KHCO\({}_{4}\) were added to the sample (0.5 mL) and then gently mixed for 15 min. The solution was then centrifuged at 10,000 rpm for 5 min at 4 \({}^{\circ}\)C to remove the precipitate. Then, 0.5 mL of 12.5% (\(w/v\)) NH\({}_{4}\) sulfamate was added to a 0.5 mL sample, which was mixed vigorously for 5 min, followed by the addition of 0.5 mL of 0.5% (\(w/v\)) 3-methyl benzothiazolinone-2- hydrazone solution. The samples were then mixed and boiled for 3 min. After cooling to room temperature, 0.5 mL of 0.83% (\(w/v\)) FeCl\({}_{3}\)\(\cdot\)6H\({}_{2}\)O was added. Keeping the sample at room temperature for 30 min. The absorbance was measured with a spectrophotometer at room temperature for 650 nm. The glucosamine content decomposed by chitin can be calculated according to the formula:  
\[\text{X}=\frac{\text{A}-A_{0}}{A_{1}-A_{0}}*10\]  
where X is the glucosamine concentration (mg/mL), \(A_{1}\) is the standard absorbance value, \(A_{0}\) is the absorbance of distilled water, A is the absorbance of the sample, and 10 is the glucosamine concentration (mg/mL). Ergosterol concentration was determined according to the method described by [PERSON] et al. (2020) [19] and [PERSON] et al. (2006) [20] with some modifications. Ultrasonic extraction of lyophilized _G. lucidum_ mycelia powder with methanol for 2 h. The supernatant was evaporated to dryness under a N\({}_{2}\) atmosphere. The residue was saponified with a 10% (\(w/v\)) KOH-75% (\(v/v\)) ethanol solution at 50 \({}^{\circ}\)C for 2 h. The mixture was subjected to hexane extraction three times, and the hexane layer was collected and evaporated to dryness under N\({}_{2}\). The residue was dissolved in methanol and subjected to high-pressure liquid chromatography (HPLC) on an Agilent 1260 (VWD/RID) apparatus equipped with a Symmetry C18 5.0 column (4.6 \(\times\) 250 mm; Waters, Milford, MA, USA). The mobile phase used the solvent of 100% methanol, and the procedure was conducted as isocratic elution with a 1.0 mL/min constant flow rate. Ergosterol content was determined and monitored at a 282 nm UV wavelength. Identification and quantification were performed according to retention time, UV spectra, and ergosterol standard (Sigma Aldrich, St. Louis, MO, USA).