Generation of Pqbp1-cKO mice

Pqbp1 gene conditional knock-out mice were generated as previously described [16]. Briefly, a target vector was generated by PCR from a BAC library (ID:RP23-404N15) and constructed using a 3.6-kb 5′ fragment containing exons 1 and 2, a neomycin resistance cassette flanked by Flp recognition target sites, a 3.9-kb fragment containing exons 3–7 between two LoxP sites, a 4.1-kb non-coding 3′ fragment, and a diphtheria toxin A gene. The target vector was introduced into ES cells (C57BL/6) using electroporation, selected with G418, and cloned. Genomic DNAs of cloned ES cells were analyzed by PCR and Southern blotting. The selected ES cells were injected into C57BL/6 blastocysts, and chimeric mice were cross-bred with C57BL/6 mice. Pqbp1-floxed mice were generated by removing the neomycin resistance cassette by cross-breeding with CAG-FLPe recombinase transgenic mice [22]. Pqbp1 conditional knockout mice were generated to cross-breed the Pqbp1-floxed heterozygous or homozygous female mice with Synapsin1-Cre transgenic heterozygous male mice [B6.Cg-Tg (Syn1-Cre) 671Jxm/J; The Jackson Laboratory, Bar Harbor, ME, USA]. Impairment of Pqbp1-cKO mice in behavioral tests such as the open field test, light-dark box, elevated plus maze test, fear conditioning test, water maze test, and rotarod test was reported in Supplementary Fig. 11 of our previous paper [16].