Fig 1: Joint pathology scores in mice and rats. A Safranin-O photomicrographs displaying a variety of OA severity and semi-quantitative scores. First score represents femoral condyle, second score is tibia; a 0, 0; b 0.5, 0; c 0.5, 2; d 2, 4; e 1, 0.5; f 2, 4; g 1, 2; h 4, 6. Among these, a, c, e, g shows the contralateral knees and b, d, f, h shows the MCP-1 intra-articulate injection knees. B The OARSI semi-quantitative scores of knees of each pairs of two subgroups at each time point; a scores of contralateral knees, despite some individuals with relatively high score (<2), most of the scores were low and stable; b scores of MCP-1 intra-articulate injection knees increased over time with some knees reaching the maximum score of 6. The rate of increase plateaued in later time-points. C HE slides of rats’ knee joints, articular cartilage was almost completely lost in both experiment and contralateral knee at 2 week after CCR2 antagonist or physiological saline injection. a The lateral tibia plateau of contralateral knee; b medial tibia plateau of experimental knee following CCR2 antagonist intra-articular injection. The cartilage degeneration score of Both A and B reached the highest grade of 5, at 2 weeks after CCR2 antagonist or physical saline injection
Fig 2: The expression of MCP-1-CCR2 axis genes and cartilage matrix markers after stimulation of MCP-1. a, b In OA chondrocytes, after stimulation of MCP-1 in culturing medium of the relative expression of MCP-1 and CCR2 mRNA increased significantly compared with unaffected OA chondrocytes; c, d the expression of Mcp-1 in OA/CHON and OA/S.F were significantly higher than RA/SF and WT/CHON. The expression of Ccr2b in OA/CHON were significantly lower than that in OA/S.F, RA/SF and WT/CHON. MCP-1 Stim, MCP-1 stimulated; RA/SF, RA synovial fibroblasts; OA/S.F, OA synovial fibroblast; OA/CHON, OA chondrocytes; WT/CHON, wild-type (normal controls) chondrocytes; RA/CHON, RA chondrocytes; and e, f, g after stimulation of MCP-1, the expression of MCP-1 and MMP13 protein increased significantly than unstimulated normal OA chondrocytes. MMP3 increased but there was not significantly higher than unstimulated normal chondrocytes. MCP-1 Stim, MCP-1 stimulated OA chondrocytes; N–N, unstimulated wild-type (normal controls) chondrocytes; N–M, MCP-1 stimulated wild-type (normal controls) chondrocytes. *P < 0.05, **P < 0.01, ***P < 0.001
Fig 3: FGF-1 treatment alleviates cell inflammation in ARPE-19 cells incubated in high-glucose medium. After pretreatment with different concentrations of FGF-1 (1, 5, and 10 ng/mL) for 2 h, ARPE-19 cells were incubated in a high-glucose (25 mM) medium for 24 h. (A) Expression levels of inflammatory mediators (ICAM-1, MCP-1, IL-1ß, and IL-6) were detected using RT-PCR. (B) Protein expression of inflammatory mediators was assessed using western blot analysis. (C) Protein expression of inflammatory mediators in cell culture medium was assessed using ELISA (n = 5 in each group). * p < 0.05, ** p < 0.01 compared with control cells incubated in low-glucose medium; # p < 0.05, ## p < 0.01 compared with ARPE-19 cells incubated in high-glucose medium. LG, low glucose; HG, high glucose; FGF-1, fibroblast growth factor type 1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.
Fig 4: Chondrocyte apoptosis assessment and the expression of CCR2 in OA chondrocytes. a MCP-1 stimulation of wild-type (normal controls) chondrocytes resulted in an increased rate in secondary necrosis. Lower left quadrant represents viable cells, the lower right quadrant are chondrocytes in an early apoptotic state, the upper right quadrant are chondrocytes in the late apoptotic state and the upper left quadrant is chondrocyte necrosis. N, unstimulated wild-type (normal controls) chondrocytes; N–M, MCP-1 stimulated wild-type (normal controls) chondrocytes; b the variance of early apoptotic state between the two subgroups was not significant and the variance in secondary necrotic rate and both was statistically significant; and c, d MCP-1 stimulation in OA chondrocytes resulted in a significant increase in the expression of CCR2. Wild-type (normal controls), unstimulated OA chondrocytes; MCP-1, MCP-1 stimulated OA chondrocytes. *P < 0.05
Fig 5: FGF-1 treatment attenuated inflammatory mediators in rats. Retinal sections were collected from 12-week-old SD rats in each group. Immunofluorescence images of inflammatory mediators (A) ICAM-1, (B) MCP-1, and (C) IL-1ß in retinal tissue. The relative densities of immunofluorescence images were quantified using ImageJ software and are presented in bar graphs (n = 4 eyes in each group). (D) Aqueous humor was collected from the rat eyeballs. ICAM-1, MCP-1, and IL-1ß protein levels were measured in each group using ELISA. Each assay was repeated three times (n = 4 eyes in each group). All data are presented as mean ± standard deviation; ** p < 0.01 compared with the control group; # p < 0.05, ## p < 0.01 compared with the diabetes group. DAPI, 4',6-diamidino-2-phenylindole; Ctrl, control group; DM, STZ-induced diabetic rat group; DM + FGF-1, fibroblast growth factor type 1 treatment group.
Supplier Page from BioLegend for LEGEND MAX(TM) Human MCP-1/CCL2 ELISA Kit