The steroid hormone 20-hydroxyecdysone binds to dopamine receptor to repress lepidopteran insect feeding and promote pupation
Fig 5
20E antagonizes dopamine function.
A. Apoptosis signal in HaEpi cells after DMSO, 20E (5 μM), dsGFP+20E (5 μM), dsDopEcR+20E (5 μM) and DA (10 μM) +20E (5 μM) treatment by using the NucView caspase-3 activity assay kit. a. The ratio of apoptotic cells (green) to the total cells (blue) in the field view was obtained. B. Proliferation signal in HaEpi cells after DMSO, 20E (5 μM), dsGFP+20E (5 μM), dsDopEcR+20E (5 μM) and DA (10 μM) +20E (5 μM) treatment by using the 5-ethynyl-2′-deoxyuridine (EdU) kit (Ribobio, Guangzhou, China). b. The ratio of proliferation cells (green) to the total cells (blue) in the field view was obtained. DAPI stained the nucleus (blue). Statistical analysis using the data from 100 × 3 cells. The yellow bars represent 100 μM. C. Western blot analysis of 20E (5 μM) or DA (10 μM) induced proteins phosphorylation for 30 min. a. b. and c. AKT-RFP-His, AKT -RFP-His, and CDK10-RFP-His. 7.5% SDS-PAGE gel. Error bars show the mean ± SD of three times’ biological repetition. ImageJ software was used to transform the image data. Significant differences were calculated by Student’s t test (*p < 0.05; **p < 0.01) or one-way analysis of variance (ANOVA, p < 0.05).