LN alterations in D6-deficient mice. (A) TPA painting leads to increased weight of D6-deficient inguinal LNs compared with WT inguinal LNs. These data were obtained by removing, and weighing, inguinal LNs from mice at rest and 24 hours after TPA painting. LNs were weighed on a ExplorerPro microbalance. (B) This weight increase is associated with an increase in D6-deficient LN cellularity. LNs were collected and dispersed into 1 mL of medium with DNAse and the cells counted on a hemocytometer. These data were obtained using WT and D6-deficient inguinal LNs collected at rest and 24 hours after TPA painting. (C) Footpad injection of LPS also increases LN cell numbers in D6-deficient, compared with WT, mice. These data are from popliteal LNs collected at rest (contralateral LNs draining PBS injected footpads), and 6 hours after LPS injection, and processed as in panel A. (D) Chemokine injection alone led to exaggerated increases in the cellularity of D6-deficient LNs. hCCL3 in PBS/0.1% BSA was injected into subcutaneously air pouches on the back of D6-deficient, or WT, mice and the cellular content of inguinal LNs, processed as in panel B, assessed 1 hour later. (A-D) Data are from 3 independent experiments with n > 3 mice per experimental group for each individual experiment.