Impaired LN development in Clec1b−/− embryos. (A) Macroscopic observation of newborns Clec1b−/−, Clec1b+/−, and Clec1b+/+ littermates revealed the absence of inguinal and mesenteric LNs in Clec1b−/−. Fraction of the number of newborn mice with detectable LNs/number of mice sacrificed. (B) Flow cytometric analysis of mesenteric LN single-cell suspensions from Clec1b−/− and Clec1b+/+ littermates newborn and E16.5 stained with CD45, CD4, ICAM-1, and VCAM-1. Percentage of CD45+ hematopoietic cells is shown in the histogram (first column); 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) Quantification of the recruitment of LTi cells (gated as shown in B second column) and of LTo cells in the CD45+ stromal fraction (gated as shown in B third column) in the LN anlage at E16.5, E18.5, and newborn stages. Data are representative of 3 to 5 embryos for each time point and phenotype across at least 2 independent experiments. Unpaired Student t test, *P < .05.