1. Utility of circulating serum miRNAs as biomarkers of early cartilage degeneration in animal models of post-traumatic osteoarthritis and inflammatory arthritis 
L.H.W. Kung, S. Zaki, V. Ravi, L. Rowley, M.M. Smith, K.M. Bell, J.F. Bateman, C.B. Little 
Osteoarthritis and Cartilage 
Volume 25, Issue 3, Pages (March 2017)
DOI: /j.joca
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

2. Fig. 1
Histopathology scoring of non-calcified femoral plus tibial articular cartilage for: chondrocyte hypertrophy/apoptosis/death (A); proteoglycan loss (B); and cartilage structural damage (C). Scatter plots displaying values for each mouse and mean (horizontal bar) ± 95% confidence intervals 1, 4 or 16 weeks after sham or DMM surgery, or mBSA immunization and intra-articular saline (Sal) or mBSA (AIA) injection (n = 6/group other than week 16 DMM n = 5). The maximum score for each parameter from two scorers blinded to treatment and time was averaged to provide the value for each individual animal. Significant differences between groups connected by lines with exact P-values for each comparison indicated above the line.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

3. Fig. 2
Serum miRNA microarray expression profiling. Scatter plots displaying normalized intensity values for each miRNA expressed above background between DMM vs sham and AIA vs Saline (SAL) at 1, 4 and 16 weeks post induction (n = 6/group other than week 16 DMM n = 5). miRNA expression was analysed by Agilent miRNA arrays and each point represents the average expression of a miRNA from 5 to 6 replicates. The trend line represents no fold change.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

4. Fig. 3
Validation of miRNA expression by qPCR. Expression of miRNAs in serum from AIA, saline (SAL), DMM and sham mice at 1 and 16 weeks was determined by qPCR. Data shown as expression relative to the expression of a housekeeper (HK), 5s rRNA. Each symbol represents an individual mouse (average of two technical replicates). Horizontal bars show the geometric mean expression. In all cases, there were no statistically significant differences between AIA vs SAL and DMM vs sham as determined by Mann–Whitney U test.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

5. Fig. 4
Comparison of miRNA expression between AIA, DMM and Sham serum samples. (A) Volcano plot showing differentially expressed miRNAs greater than 1.5 fold change and adj. P-value <0.05 at 1 week AIA vs DMM. 71 miRNAs were significantly dysregulated (P < 0.05) in 1 week AIA vs DMM (highlighted in green + blue + red). Of those, 36 miRNAs were up-regulated >1.5 fold with P < 0.05 (highlighted in red) and 13 miRNAs were down-regulated >1.5 fold with P < 0.05 (highlighted in blue) in 1 week AIA vs DMM (B) qPCR analysis directly comparing miRNA expression of AIA and DMM samples at 1 week post induction. Data shown as expression relative to the expression of a housekeeper (HK), 5s rRNA. Each symbol represents an individual mouse (average of two technical replicates). Horizontal bars show the geometric mean expression. *P < 0.05, **P < 0.01 in AIA vs DMM as determined by Mann–Whitney U test. (C) Volcano plot showing differentially expressed miRNAs greater than 1.5 fold change and adj. P-value <0.05 at 1 week AIA vs Sham. 49 miRNAs were significantly dysregulated (P < 0.05) in 1 week AIA vs Sham (highlighted in green + blue + red). Of those, 20 miRNAs were up-regulated >1.5 fold with P < 0.05 (highlighted in red) and 9 miRNAs were down-regulated >1.5 fold with P < 0.05 (highlighted in blue) in 1 week AIA vs Sham. (D) Venn diagram comparing the degree of overlap between AIA vs DMM and AIA vs Sham.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

6. Fig. 5
Normalized array data showing expression levels of miRNAs in 1 week AIA, saline (SAL), DMM and sham. Plots demonstrate subtle non-significant changes in miRNA expression in AIA vs SAL, DMM vs sham and AIA vs sham. Statistically significant, dynamic changes were observed between AIA and DMM models. Differences between groups connected by lines with exact P-values for each comparison indicated above the line. Each symbol represents an individual replicate. Horizontal bars show the mean expression of the group.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions