Fig. 5
From: Control of Treg cell homeostasis and immune equilibrium by Lkb1 in dendritic cells
Lkb1 restrains NF-κB signalling to inhibit Ox40l expression. a Immunoblot analysis of total and phosphorylated p65, IKKα and IκBα expression in WT and Lkb1-deficient DCs treated or untreated with LPS (1μgml−1) for 40 min. b Immunoblot analysis of p65 protein in the nucleus of splenic DCs from Lkb1f/f and Cd11cCreLkb1f/f mice. c Real-time PCR analysis of the mRNA level of Ox40l in splenic DCs from Lkb1f/f and Cd11cCreLkb1f/f mice treated with or without an NF-κB inhibitor (SC75741) for 3 days. d, e Flow cytometric analysis (d) and quantification (e) of relative MFI of OX40L expression on DCs in LNs from Lkb1f/f and Cd11cCreLkb1f/f mice treated or untreated with an NF-κB inhibitor (SC75741) for 3 days. f, g Flow cytometric analysis (f), percentage and number (g) of CFSE-labelled Treg cells co-cultured with DCs from Lkb1f/f and Cd11cCreLkb1f/f mice treated with or without SC75741 for 4 days. h, i Flow cytometric analysis (h), percentage and number (i) of CFSE-labelled Treg cells (CD4+Foxp3+, 2×106) sorted from B6 mice transferred together with or without DCs (1×106) sorted from Lkb1f/f or Cd11cCreLkb1f/f mice treated with or without SC75741 into irradiated NSG mice via tail vein injection for 3 days. j Relative enrichment of NF-κB p65 to Ox40l promoter in Lkb1f/f or Cd11cCreLkb1f/f DCs treated with LPS (1 μg ml−1) for 3 h, determined by ChIP. The results are presented as the mean ± S.E.M., *P < 0.05, **P < 0.01, ***P < 0.001, by Student's t-test (c, e, g, i). Data are pooled from e, g, i, j or are representative of at least three a, b, d, f, h independent experiment with similar results
