Fig. 6: Increased [Ca²⁺]_ex-induced, CPP-dependent inflammasome activation in RA.

a–c Detection of IL-1β a, IL-1α b, and IL-18 c in cell culture supernatants of freshly isolated peripheral blood monocytes from patients with RA (RA), psoriatic arthritis (PsA), systemic lupus erythematodes (SLE), or healthy donors (HD) after 16 h of incubation with LPS and [Ca²⁺] in RPMI1640/10%FBS/5.6 mM [P_i] media. *Indicates level of significance for comparison against the same concentration in healthy donors; ^#for comparison between RA with and without DMARD. Comparison to RA without DMARD in panel a: p = 0.0073 (1.2 mM) and p < 0.0001 (1.7 mM) for PsA, p = 0.0013 (1.2 mM) and p = 0.0094 (1.7 mM) for SLE. d Stimulation of freshly isolated peripheral blood monocytes from RA patients and healthy donors (HD) with 2x CPPs and the indicated added [Ca²⁺] in 1 mM [P_i] RPMI1640/10%FBS. e Stimulation of human monocyte-derived macrophages from RA patients (RA) or healthy donors (HD) with the indicated added [Ca²⁺] and detection of IL-1β in cell culture supernatants after 16 h of incubation in 5.6 mM [P_i] RPMI1640/10%FBS; f Calcein-CPP uptake of human monocytes from either RA patients (RA) or healthy donors (HD) after stimulation with 2.5 mM added [Ca²⁺] for 45 min, detection and quantification was done by imaging flow cytometry (ImageStream^XMarkII); g Representative Western blot and dot plot showing expression levels of CaSR in freshly isolated monocytes from either healthy donors (HD) or RA patients (RA), Ponceau staining of PVDF membranes was used for quantification of CaSR protein (see Supplementary Fig. 3c). Box-and-whisker plots show median, 25–75th percentile, and min/max whiskers. Patient numbers are indicated in the figure. Statistical analysis was performed using two-tailed Mann–Whitney U test. p-values are indicated as *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.