1. Leptin promotes apoptosis and inhibits autophagy of chondrocytes through upregulating lysyl oxidase-like 3 during osteoarthritis pathogenesis 
Z.M. Huang, S.H. Du, L.G. Huang, J.H. Li, L. Xiao, P. Tong 
Osteoarthritis and Cartilage 
Volume 24, Issue 7, Pages (July 2016)
DOI: /j.joca
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

2. Fig. 1
LOXL3 mRNA was positively correlated with leptin concentration. (A) LOXL3 expression was significantly higher in osteoarthritis cartilage (n = 33) than in healthy cartilage (n = 7) from GSE57218 dataset. (B) LOXL3 mRNA level of cartilage from osteoarthritis patients (n = 45) and health volunteers (n = 25) was evaluated by real-time PCR. (C) Leptin concentrations in joint SF from osteoarthritis patients (n = 45) and health volunteers (n = 25) were measured by using leptin radioimmunoassay kit. (D) Pearson correlation analysis between LOXL3 and leptin (P < 0.0001, n = 45).
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

3. Fig. 2
Function analysis of LOXL3 in a rat osteoarthritis model. Eighteen rats were randomly divided into three groups: Sham: sham-operation, ACLT+vehicle: ACLT model injected with vehicle and ACLT+RAPA: ACLT model injected with rapamycin. Ten weeks after surgery, the cartilage and joint SF samples were collected. (A) After fixation, decalcification and embedding, sections were cut and stained with Safranin-O-fast green. Scale bar: 100 μm. (B) LOXL3 mRNA level in rat cartilage were evaluated by real-time PCR (n = 6). (C) SF leptin concentrations were assessed by radioimmunoassay (n = 6). (D) Positive correlation between LOXL3 and leptin as determined by Spearman correlation analysis (P < 0.0001, n = 18). (E, F) The expression of cleaved caspase 3, Bcl-2, LC3 and Beclin1 were measured by western blot. The left panel showed representative western blot and the right panel showed the quantification of western blot (n = 3). *P < 0.05, **P < 0.01 vs sham group; #P < 0.05, ##P < 0.01 vs ACLT+ vehicle group.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

4. Fig. 3
Silencing of LOXL3 promoted the proliferation and inhibited the apoptosis of chondrocytes. (A) Primary cultured chondrocytes were treated with a serial dosages of leptin for 48 h. LOXL3 protein level was detected by western blot (n = 3). (B) Primary chondrocytes isolated from rat knee joint with ACLT surgery were divided into three groups: Control: ACLT+leptin (20 ng/mL), siNC: ACLT+leptin (20 ng/mL) + siNC, and siLOXL3: ACLT+leptin (20 ng/mL) + siLOXL3. The chondrocytes viability as tested by CCK-8 (n = 3). (C) Detection of cell apoptosis by Annexin V-PI staining and flow cytometry analysis (n = 3). (D) Western blot analysis of LOXL3 and apoptosis associated protein expression. **P < 0.01, ***P < 0.001, vs siNC group (n = 3).
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

5. Fig. 4
LOXL3 inhibited the autophagy of chondrocytes via activating mTORC1. (A)Western blot analysis of Beclin1 and LC3 (n = 3). Control: ACLT+leptin (20 ng/mL), siNC: ACLT+leptin (20 ng/mL) + siNC, and siLOXL3: ACLT+leptin (20 ng/mL) + siLOXL3. **P < 0.01 vs siNC group. (B, C) Primary chondrocytes isolated from rat knee joint with ACLT surgery were divided into four groups: NC: cells treated with NC virus, LOXL3: cells treated with LOXL3 virus, RAPA: cells treated with rapamycin, and LOXL3+RAPA: cells treated with LOXL3 virus and rapamycin. Western blot analysis of Beclin1, LC3, p-p70S6K, p70S6K, p-AKT and AKT. *P < 0.05, **P < 0.01 vs ACLT+NC group; #P < 0.05, ##P < 0.01 vs ACLT+LOXL3 group (n = 3).
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions