After eclosion, the 3-day-old females were anesthetized on ice and a total of 1 μL agomiR/antagomiR solution (20 μM) was injected into the ventral abdomen of female adults using an Eppendorf micromanipulation system (FemtoJet 5247; Eppendorf, Hamburg, Germany) as described previously [65 (link)]. The same volume of random shuffled sequence was used as a negative control (NC). At 24 h post injection, flies were sacrificed to dissect the ovary and fat body for RNA extraction and the subsequent qRT–PCR analysis (10 flies in each treatment group). The miRNA mimic (agomiR), inhibitors (antagomiR) and negative control (NC) oligonucleotides were purchased from GenePharma (Shanghai, China). The sequences are listed in Table S2.
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Ovary and Fat Body miRNA Analysis
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Abdomen
Adults female
Antagomir
Fat body
Females
Flies
Inhibitors
Micromanipulation
Mirna
Oligonucleotides
Ovary
Corresponding Organization : Huazhong Agricultural University
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Variable analysis
independent variables
- Injection of agomiR/antagomiR solution (20 μM, 1 μL) into the ventral abdomen of female adults
- MiRNA expression in the ovary and fat body as measured by qRT-PCR
- Injection of random shuffled sequence (1 μL) as a negative control (NC)
- Negative control (NC): Injection of random shuffled sequence
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