The exosomes were added into 1 mL Trizol (Invitrogen, USA), blown uniformly with RNA‐free gun‐head repeatedly, and then placed at room temperature for 5 minutes. A total of 200 μL chloroform was added and incubated at room temperature for 10 minutes with severe shock for 15 seconds. Then, the exosomes were centrifuged at 12 000 g and 4°C for 15 minutes and 400 μL supernatant collected. Additional 400 μL isopropanol (Sangon Biotech) was added, mixed well, and left to dry at room temperature for 10 minutes. Afterwards, the mixture was centrifuged at 12 000 g and 4°C for 10 minutes and the supernatant discarded. Again, 1 mL 75% ethanol was added, gently washed and precipitated, centrifuged it at 4°C for 5 minutes and the supernatant discarded. After drying at room temperature, an appropriate amount of DEPC water was added to dissolve and kept on ice for the next experiments.

Follicular fluid samples from ten OHSS patients and ten normal controls were used for exosomal RNA sequencing. Library preparation and sequencing were conducted at Anoroad. A total of 1 μg total RNA per sample was used for the small RNA library. After the total RNA samples were quantified, the RNA fragments were fractionated on a 15% polyacrylamide gel (Invitrogen) and small RNAs ranging between 15 and 30 nucleotides (nt) were used for library preparation. The two ends of the separated RNA fragments were connected respectively. Small RNAs were reverse transcribed by RT primers and amplified by PCR. The PCR products were sequenced using the Illumina HiSeq 2500 platform (Illumina Inc).

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