Generation and Genotyping of miR-132/212 KO Mice

The generation of miR-132/212 KO mice has been described previously (15, 16). Briefly, the floxed miR-212-miR132 mice were generated by gene targeting in an ES cell line from C57BL/6N, then crossed with a Cre delete line to remove miR-212-miR-132 region. The result miR-212-132^−/− line is in C57BL/6J. The animal experiment was all carried out using this strain with age and sex-matched C57BL/6J mice as wildtype (WT) control. For genotyping, genomic DNA was extracted from ear clippings using the genomic DNA isolation kit (Sigma, Cat. XNATS). PCR was done with the GC-Rich PCR kit (TAKARA, Cat. RR002C) with miR-132/212 primers, as shown in Supplementary Material. PCR products were separated on 1% agarose gel: WT gave a band at 1,076 bp and the KO at 392 bp.