Ago2-dependent miRNAs in Drd2 neurons are induced by cocaine and regulate genes involved in cocaine addiction. (A) Ago2-regulated miRNAs in Drd2 neurons are induced by cocaine. The Venn diagram shows the number of miRNAs that are down-regulated (>1.5 fold) in the striatum of Camk2a-Cre; Ago2^fl/fl mice (blue; n = 6/genotype) and miRNAs that are up-regulated (>1.45-fold) in Drd2 neurons in response to acute cocaine as compared with saline injections (red; n = 3/genotype). The overlapping miRNAs are shown in yellow. The bar graph displays all 23 miRNAs that are up-regulated in response to acute cocaine in Drd2 neurons (blue bars) and down-regulated in the absence of Ago2 in the striatum (red bars). RQ of each miRNA was calculated using the ΔΔCt method; a maximum standard deviation for the ΔCt values of 0.8 was used as a cut-off for each miRNA. (B) Ago2-dependent, induced by cocaine and Drd2-enriched (ADICD) miRNAs regulate genes important in cocaine addiction. The bar graph displays the average percentage of luciferase expression of FosB-, Mef2d-, and Cdk5r1- 3′ UTR-luciferase vectors in the presence of miR-369-3p, miR-324-5p, or a scrambled control as compared with their respective vector controls. Each sample was analyzed in triplicates, and three independent experiments were performed. Statistical analysis was performed using Student’s t test. Error bars indicate + SEM. *, P < 0.05; **, P < 0.01.