Decreased NF-κB regulated gene transcription in patient fibroblasts. (A) qPCR gene expression using TNF-treated fibroblasts from three controls (C) and two patients (Pts). (B) Heat map with unsupervised clustering showing the top 5% of genes with the most difference in TNF-treated fibroblasts from three controls and two patients, as determined by RNA-seq in one experiment. (C) Log₂-fold change in the expression of indicated genes using RNA-seq data from two patients relative to three controls. (D) qPCR analysis of WT RELA expression in GFP⁺ fibroblasts from two controls and two patients after nucleofection with a vector encoding GFP or GFP-RELA. (E–G) RT-qPCR analysis of the expression of BCLA21 and IL6 in GFP⁺ fibroblasts from two controls and two patients after nucleofection with a vector encoding GFP or GFP-RELA and stimulated as indicated. Gene expression data in A and D–G were normalized to β-glucuronidase, expressed as the fold increase relative to unstimulated cells and were pooled from at least two independent experiments. Columns and bars represent means ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; n.s., not significant.