Fig. 1

Characterization of the circRIMS2/miR-3968 pathway in AD models. a Volcano plots displaying 116 differentially expressed circRNAs and 29 differentially expressed miRNAs in the hippocampus of 4-month-old wild-type (WT) and APP/PS1 mice by sequencing (n = 3). circRIMS2 was increased while miR-3968 was decreased. The cut-off P value was 0.05. b qRT-PCR analysis of dysregulated ceRNA pairs involving 11 circRNAs (left, n = 4) and 8 miRNAs (right, n = 3) in the hippocampus of 4-month-old APP/PS1 mice. c qRT-PCR analysis of circRIMS2 and miR-3968 levels in the hippocampus of 4-month-old 3 × Tg and WT mice (n = 4). d qRT-PCR analysis of circRIMS2 and miR-3968 levels in scramble- and Aβ-treated mouse primary cortical neurons (n = 4). e Relative expression of circRIMS2 (left) and miR-3968 (right) in the hippocampus of APP/PS1 and WT mice at diverse stages (n = 4). f The distribution of circRIMS2 and miR-3968 in N2a cells was detected by FISH. CircRIMS2 and miR-3968 were labeled in red and green, respectively; nuclei were labeled with DAPI (blue). Scale bars, 50 μm. g Schematic illustration of circRIMS2 formation through the circularization of exon 4 and 19 in RIMS2. The back-splicing junction site of circRIMS2 was validated by Sanger sequencing. h PCR analysis of circRIMS2 and GAPDH in cDNA and genomic DNA (gDNA) amplified by convergent and divergent primers. i PCR analysis of circRIMS2 and RIMS2 from 3 μg RNA treated with 10U RNase R (GENESEED, Guangzhou, China) for 10 min. j qRT-PCR detected the abundance of circRIMS2 and RIMS2 in N2a cells treated with ActD (2 μg/ml) for 4 h (n = 3). k qRT-PCR analysis of circRIMS2 in the nucleus (Nuc) and cytoplasm (Cyt) of N2a cells (n = 3). Anti-Ago2 RNA immunoprecipitation was performed in N2a cells, and circRIMS2 was enriched by Ago2 (n = 4). Data are presented as mean ± S.E.M., and two-tailed t-tests were used unless otherwise specified. *P < 0.05, **P < 0.01, ***P < 0.001