Increased expression of p15Ink4b mRNA in cDCs. (A) FACS plots depicting the gating strategy for FACS sorting of mouse FACS-sorted c-Kit⁺ cells from lineage-negative enriched BM progenitors (left) and CD11c⁺/CD11b⁺ splenic cDCs from enriched splenic DCs (right). DCs were enriched using a negative-enrichment cocktail containing biotinylated Abs against murine CD2, CD90.2, CD19, Ly-6G, and TER-119. (C) CD11c⁺CD11b⁺ BM-DCs from the GM-CSF and IL-4 BM cultures. (B,D,G) Real-time qPCR analysis of p15Ink4b mRNA levels of the sorted populations of cells. (E) Human CD34⁺ progenitors and CD34-derived CD1c⁺CD11b⁺ cDCs. Cells were harvested at the indicated time points. (F) Bright-field images of human CD34⁺ progenitors (left) and CD34-derived DCs on day 5 of culture (right). Scale bar represents 20 μm. Images were collected using an Olympus iX51 inverted microscope, a EXi Aqua Camera (QImaging), and the acquisition software IPlab Version 4.08 (BD Biosciences), and were processed in Adobe Photoshop Version 5.0.