Modulation of NF-κB response genes by SUMOylation-deficient WASp mutant. (A) The effect of SUMOylation-deficient WASp mutants (PSM and V75M) on mRNA expression levels of multiple NF-κB response genes in human TH1-skewed cells, in the presence (+) or absence (−) of panHDAC inhibitors (HDACi). Shown are some representative genes from the different gene ontology (GO) categories selected from the complete dataset including their heatmaps displayed in supplemental Figure 5. The mRNA expression, quantified by RT-PCR array, in SUMOylation-deficient WASp-expressing TH1 cells was compared with that in FL-WASp-expressing TH1 cells after normalizing to their respective control genes (ACTB and RPLP0) and then represented as fold upregulation (activated) or downregulation (repressed). (B) Flow cytometry showing histogram profiles for intracellular expression of indicated cytokines and transcription factors (after permeabilization) and cell surface expression of TLR1 (without permeabilization) along with their corresponding geometric means (GMs). Staining with isotype IgAb (gray histogram) is shown as a control. NEM+, treated with NEM.
