Fig 2: Signaling output of different innate immune stimuli in patient-derived cells AFibroblasts were stimulated with 1 µg/ml LPS for the indicated times and subjected to analysis by Western blot. One representative out of three independent experiments is shown.BFibroblasts were stimulated with LPS for 24 h. Concentration of IL-6 in the supernatant was determined by ELISA. Data are presented as mean ± SD of three individual experiments performed in two technical replicates; *P = 0.012; multiple t-tests corrected for multiple comparisons using the Holm–Sidak method.CB cells were treated with 100 ng/ml CD40L for the indicated times and subjected to analysis by Western blot. One representative out of three independent experiments is shown.DFibroblasts were incubated with 50 µg/ml cycloheximide (CHX) for the indicated times and subjected to analysis by western blot. One representative (left panel) out of 4 individual experiments analyzed by densitometry (right panel) is shown. Data are presented as mean ± SD. *P = 0.02, unpaired t-test.E, FFibroblasts (E) or B cells (F) were treated with 0.1 µg/ml TNF, 0.1 µg/ml TRAIL, 100 µg/ml poly(I:C), 1 µg/ml LPS or left untreated, and viability was determined after 24 h. Data are presented as mean ± SD of three independent experiments; dots represent individual experiments performed in three technical replicates; ns, non-significant, unpaired t-test.G, HFibroblasts were incubated with 50 µg/ml cycloheximide (CHX) or DMSO and stimulated with the indicated concentrations of TRAIL (G) or poly(I:C) (H). Viability was measured after 24 h. Data are presented as mean ± SD of three individual experiments performed in three technical replicates. Source data are available online for this figure.

Fig 3: Expression profile of cytolytic enzyme (granzyme B) and functional cytokines (IFN?, TNF) in CD8+ T cells during decitabine treatment at day 5 post-stimulation(A–C) (A) MFI expression of Granzyme B. Functional cytokines are represented as frequency for (B) IFN?+ and (C) TNF+ expressing T cells. Figure reprinted with permission from Loo Yau et al. (2021).

Fig 4: Recommended 96-well set-up for restimulation to detect functional cytokines TNF and IFN?Cocktail 1: Cell Stimulation plus Protein secretion inhibitor cocktail. Cocktail 2: Protein secretion inhibitor cocktail. Image made with BioRender.

Fig 5: The patient’s inflammatory plasma protein profile is converted to normal after treatment with the anti-TNF therapeutic Adalimumab Plasma was collected from the patient before and after start of therapy with the anti-TNF therapeutic Adalimumab. Plasma from two healthy controls was used as comparison. The Proteome Profiler Human Cytokine Array was performed according to the manufacturer’s instructions.The concentration of Adalimumab in the patient’s serum is shown (target concentration: > 5 µg/ml).