Normal LN development in Clec1bfl/flPF4-Cre newborns. (A) Macroscopic observation of newborns Clec1bfl/flPF4-Cre and Clec1bfl/fl littermates revealed the presence of inguinal and mesenteric LNs in Clec1bfl/flPF4-Cre newborns. Fraction of number of newborn mice with detectable LN/number of mice sacrificed. (B) Flow cytometric analysis of mesenteric LN single-cell suspensions from Clec1bfl/flPF4-Cre and Clec1bfl/fl newborn littermates stained with CD45, CD4, ICAM-1, and VCAM-1. Percentage of CD45+ hematopoietic cells is shown in the histogram; percentages of CD45+CD4+ LTi cells (second column) and of ICAM-1highVAM-1high LTo cells in the CD45− stromal fraction (third column) are shown in the dot plots. (C) Immunofluorescence staining of sections from newborn mesenteric LNs showing the CD31+ blood endothelial vasculature in red and the Lyve-1+ LECs in green forming the subcapsular sinus (first column) and the Gp38+ stromal cell compartment in red and CD4+ cells in blue (second column) in Clec1bfl/fl (upper row) and Clec1bfl/flPF4-Cre littermates (lower row).