Agilent vials

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)].

N. benthamiana leaves were collected 6 days after infiltration
and immediately frozen in liquid nitrogen. Samples were lyophilized
(CHRIST Alpha 1-4 LD Plus, Martin Christ, Osterode am Harz, Germany)
and powdered with a ball mill grinder (MM301, RETSCH, Haan, Germany)
for 2 × 1 min at 29 Hz. Lyophilized samples were weighed with
a precision scale, and 100 mg of each was suspended in 5 mL of 80%
(v/v) aqueous methanol and sonicated for 20 min, followed by centrifugation
for 15 min at 3000 rpm. Two methanol extractions were performed for
each sample, and supernatants were combined into 50 mL Falcon tubes
and evaporated to dryness (SpeedVac SPD 300DDA, Thermo Scientific,
Bellefonte, USA).
Crude plant extract samples were mixed with
500 μL of 10 mM (MES)-KOH (2-(N-morpholino)
ethanesulfonic acid) and 150 μL of 5 mg/mL almond β-glycosidase
(Sigma-Aldrich, St. Louis, USA) and incubated at 37 °C for 15
h with gentle shaking. Samples were then transferred into glass KIMAX
tubes, and raspberry ketone was extracted with 4 mL of methyl tert-butyl ether (MTBE), transferred into 2 mL Agilent vials,
and evaporated to dryness under nitrogen flow.

Samples of yeast cultivations
were collected on seven separate days. From each sample, 1 mL was
collected into 2 mL Eppendorf tubes and centrifuged at 13 000
rpm to separate the cell mass. The liquid fraction was transferred
to a new 2 mL Eppendorf tube and was stored at −20 °C
prior to extraction. Extraction of raspberry ketone was carried out
in glass KIMAX tubes with 2 mL of MTBE. Samples were agitated with
cross bar magnets for 20 min on a magnetic stirrer and centrifuged
for 5 min at 3000 rpm with a KIMAX centrifuge, and the supernatant
was transferred into 2 mL Agilent vials and evaporated until dry under
nitrogen flow.