Primers for mir 30b and u5 u6

Total RNA was extracted from hippocampal tissues or cells using the RNeasy Mini Kit and treated with RNase-free DNase as described previously (Gao et al., 2022 (link); Song et al., 2019 (link); Zhang et al., 2014 (link)). 1 μg of total RNA was used with 4 μl 5× iscript reaction mix and 1 μl iscript reverse transcriptase. Samples (20 μl) were incubated for 5 min at 25 °C, and then were then heated to 42 °C for 30 min. The reaction was stopped by heating to 85 °C for 5 min. The primers for miR-30b and U5/U6 were obtained from QIAGEN (Valencia, CA). The primers for glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were selected using Beacon Designer Software (BioRad) and synthesized by IDT (Coralville, IA): forward primer, reverse primer (amplicon size), genebank number: 5’-ACCACAGTCCATGCCATCAC-3’, 5’-ACCTTGCCCACAGCCTTG-3’ (134 bp), M32599. All the PCR products were verified by sequencing. We used the relative CT method to calculate RNAs as the fold increase or decrease, which was determined relative to the control after normalizing to a housekeeping gene using 2^−ΔΔCT, where ΔCT is (gene of interest CT) - (GAPDH CT), and ΔΔCT is (ΔCT treated) - (ΔCT control), as described previously (Sang et al., 2005 (link); Zhang and Chen, 2008 (link); Zhang et al., 2014 (link); Zhang et al., 2015a (link)).