Fig. 1

IL-1β induces CXCR3 expression on MSCs. a Immunofluorescence staining of CXCR3 (green) in control MSCs and MSCs stimulated with 100 ng/ml IL-1β at 15, 30, and 180 min; cellular nuclei were stained in blue. Negative control: CXCR3 antibody without secondary antibody. Scale bar: 50 μm. b Quantitative fluorescence intensity results analyzed by Image J; 10 cells were quantitated in each experiments of control MSCs and MSCs stimulated with 100 ng/ml IL-1β at 15, 30, and 180 min. The data represent mean ± SD (n = 3). Statistical analysis was determined by Student’s t test and one-way ANOVA. *P < 0.05, **P < 0.01. c MSCs were stimulated with 100 ng/ml IL-1β at 15, 30, and 180 min. The cytosolic and membrane protein were fractionated by Mem-PER™ Plus Membrane Protein Extraction Kit, then the expression of CXCR3 (41 kDa) protein was detected by Western blotting. β-actin was used as a cytosolic marker whereas pan-cadherin was used as a cell membrane marker. d Quantitative results of the Western blotting of CXCR3 cytosolic and membrane protein expression of c. The data represent mean ± SD (n = 3). Statistical analysis was determined by Student’s t test and one-way ANOVA. *P < 0.05 versus control, ***P < 0.001 versus control. e MSCs were pretreated with protein synthesis inhibitor, cycloheximide, at concentration of 20 μg/ml, then stimulated with IL-1β for 30 min. The expression of CXCR3 membrane protein was detected by Western blotting. f Quantitative results of the Western blotting of CXCR3 membrane protein expression of e. The data represent mean ± SD (n = 3). Statistical analysis was determined by Student’s t test and one-way ANOVA. *P < 0.05