Figure 1: Mbd2 regulates expression of several key DC pathways associated with APC function.

(a) WT or Mbd2^−/− BMDCs cultured on multichamber glass slides were stained with phalloidin (green) and 4,6-diamidino-2-phenylindole (blue) and surface area analysed by confocal microscopy. Photomicrographs are representative images from five fields in one experiment of three (scale bars, 38 μM (top panel) and 7 μM (bottom panel)). CD11c staining on WT (blue) and Mbd2^−/− (red) BMDCs (one of six experiments) was assessed by flow cytometry. (b) Heat map showing the mRNA signature of WT versus Mbd2^−/− BMDCs (119 genes, log2 normalized intensity, twofold change-filtered, P<0.05 (moderated t-test), three biological replicates per genotype). (c) Heat map showing 18 downregulated Mbd2^−/− versus WT BMDCs genes selected based on putative function following network analysis. (d) Heat map showing nine upregulated Mbd2^−/− versus WT BMDCs genes selected based on putative function following network analysis. (e) To validate microarray data, mRNA expression of genes of interest were assessed by qPCR (normalized against Hprt, a.u.), surface protein expression measured by flow cytometry and secreted protein by ELISA, comparing WT and Mbd2^−/− BMDCs. Results are mean+s.e.m. (three replicate wells, one of at least six experiments). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 (Student’s t-test). a.u., arbitrary units; gMFI, geometric mean fluorescence intensity.