Enhanced PI3K-PKB activity improves CD34-derived pDC development. (A) CD34⁺ HPCs were differentiated toward pDCs in the presence or absence of VO-OHpic. After 2 weeks, cells were counted with trypan blue exclusion and analyzed for the expression of CD123, BDCA-2, and BDCA-4 by flow cytometry. Absolute numbers of pDCs per well were calculated, and the fold increase of pDCs in inhibitor cultures compared with control cultures was determined. Representative FACS plots showing pDC percentages and mean ± SEM absolute pDC numbers are shown (n = 6). (B) CD34⁺ HPCs, retrovirally transduced with myrPKB or a control vector, were differentiated toward pDCs. The percentage of eGFP⁺ cells was analyzed 2 days after transduction to determine the transduction efficiency. After 2 weeks, cells were counted with trypan blue exclusion and analyzed for eGFP, CD123, BDCA-2, and BDCA-4 by flow cytometry. Absolute numbers of eGFP⁺ pDCs per well were determined and corrected for the difference in transduction efficiency. The fold increase of pDCs in myrPKB cultures compared with control cultures was determined. Representative FACS plots showing pDCs within the eGFP⁺ population and mean ± SEM absolute eGFP⁺ pDC numbers are shown (n = 6). *P < .05, Wilcoxon signed rank test.