Impaired development of miR-142–deficient CD4⁺ DCs in vitro and enrichment of miR-142 target expression in miR-142–deficient DCs. (A) Flow cytometric analysis of BM cells of miR-142^−/− mice and WT littermates cultured for 6 days in presence of 200 ng/mL of FLT3-L quantifying percentages of CD4⁺ and CD8α⁺ DC equivalents. CD4⁺ DC equivalents were identified as PDCA-1⁻CD11c⁺CD24^intCD172a⁺CD11b^high cells and CD8α⁺ DC equivalents were identified as PDCA-1⁻CD11c⁺CD24^highCD172a⁻CD11b^int cells. Each symbol represents BM cells derived from independent mice. One representative experiment of 2 is shown. (B) Graphic summary of data. *P < .05 was considered significant using a Student 2-tailed t test. (C) Sorting of in vitro FLT3-L–cultured CD4⁺ and CD8α⁺ DC equivalents generated from miR-142^−/− mice and WT littermates. Notice the strong reduction of CD172⁺ DCs in miR-142^−/− culture. (D) Heat map depicting expression of genes showing at least a 2-fold expression difference in 1 of the 4 cell populations tested. Clustering was performed using the Pearson correlation as the distance metric. Intensities of red and blue indicate increased or decreased mRNA levels, respectively. (E) Statistical analysis showing the P values for the enrichment of miR-142-3p and miR-142-5p targets within the 5 detected clusters. Predicted and conserved targets taken from TargetScan, P values were calculated using the hypergeometric test. Black color indicates a P value of 1, gray a P value of < 1, and red a statistically significant enrichment with P < .05.