2.5 Compound eyes sampling, RNA, and protein extraction
After the exposure experiment, the fresh compound eye mixture of 3O. oratoria s in each tank were used for mRNA extraction, and the compound eye mixture of the remaining 3 O. oratoria s were used for total protein extraction. The mRNA extraction processes were as follows: Liquid nitrogen is added to the individual sample and fully ground, and the high-quality total RNA (The RNA integrity number detected by Agilent 2100 bioanalyzer is greater than 7) was extracted using the standard Trizol Reagent Kit (Sangon Biotech). Subsequently, we use RNA purification Beads (Illumina) to adsorb the mRNA from total RNA and then use Beads Binding Buffer (Illumina) to clean the adsorbed mRNA three times. The eluted mRNA was incubated at 94°C for 8 minutes to achieve mRNA purification. The total protein extraction processes were as follows: SDS lysate (Macklin) was added to the individual sample that were fully ground with liquid nitrogen to promote complete sample cleavage. PMSF (ThermoFisher) was added to the lysed sample to inhibit protein hydrolysis. The solution was ultrasonically broken 3 times on ice, and the total protein was then obtained in the supernatant by centrifugation.