Secreted Wnt-1 and Jagged-1 block complete MDDC differentiation. Flow cytometric analysis showing DC-SIGN and CD14 expression of MDDCs differentiated with IL-4 and GM-CSF plus (A) CM from cells transfected with empty pcDNA3 vector (Mock), WNT1, JAG1, or a combination of WNT1 and JAG1 (WNT1 + JAG1) expression vectors. Top row of dot plots: MDDCs differentiated with IL-4 and GM-CSF plus CM from untransfected cells (Media control). Quadrants were set based on isotype controls for each time point. (B) Flow cytometric data were quantified for comparison of DC-SIGN/CD14 expression ratios at day 3 in differentiating MDDCs. Ratios were calculated using MFI of DC-SIGN/ MFI of CD14 and normalized to media control. (C) Flow cytometry analysis showing DC-SIGN and CD14 expression of MDDCs differentiated with IL-4 and GM-CSF plus secondary anti-Fc antibodies only, preclustered ephrinB2-Fc, or preclustered Jagged-1-Fc. Quadrants were set based on isotype controls for each time point. (D) DC-SIGN/CD14 expression ratios at day 3 were calculated using MFI of DC-SIGN/ MFI of CD14 and normalized to media control.