Fig. 5: Calcium-macrophages have a pro-inflammatory and invasive phenotype.

a–g Bar charts of cytokines detected in the supernatants of macrophages stimulated with 10 ng/ml LPS or LPS and 2.5 mM calcium for 24 h (a–e) or incubated without additional stimuli (f, g). Macrophages were differentiated from monocytes from healthy donors (HD, n = 4) or RA patients (RA, n = 9). h PGE2 detected in the supernatants of macrophages stimulated with 10 ng/ml LPS for 24 h. Macrophages were differentiated from monocytes from healthy donors (HD, n = 4) or RA patients (RA, n = 9). i COX-2 expression in resting macrophages or macrophages stimulated with 10 ng/ml LPS for 24 h (n = 4). Shown is one representative western blot and quantification of COX-2 expression. j–m Scratch assay of macrophage monolayers (n = 9). j Representative phase contrast images taken with IncuCyte. Purple line indicates the mask for initial scratch width. Magnification 10x. k Change of relative wound density during 38 h after scratch. l Area under the curve (AOC) of relative wound density. m Area under the curve (AOC) of relative wound density of macrophages from healthy donors (n = 9) and RA patients (n = 9). a–i, l, m Bar charts show mean ± s.e.m. Statistical analysis was performed using two-tailed t test. Significance levels are given or marked with * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 (comparisons between LPS-stimulated macrophages and calcium-macrophages) or # p ≤ 0.05, ## p ≤ 0.01, ### p ≤ 0.001 (comparisons between LPS/calcium-stimulated macrophages and calcium-macrophages).