Journal: Aging (Albany NY)

Article Title: Cabozantinib prevents AGEs-induced degradation of type 2 collagen and aggrecan in human chondrocytes

doi: 10.18632/aging.205186

Figure Lengend Snippet: Cabozantinib increased type II collagen and aggrecan levels in AGEs-treated SW1353 chondrocytes. SW1353 chondrocytes were stimulated with 100 μg/ml AGEs with or without 10 and 20 μM Cabozantinib for 24 h. The type II collagen and Aggrecan levels were detected by the Western blotting assay (n=6, *, ** P <0.05, 0.01 vs. Control group, #, ## P <0.05, 0.01 vs. AGEs group).

Article Snippet: After separation using a 12% sodium dodecyl sulfate (SDS)-polyacrylamide gel (PAGE), proteins were moved to the polyvinylidene fluoride (PVDF) membrane, followed by loading with primary antibodies against type II collagen (1:800, Cat#28459-1-AP, Proteintech, USA), Aggrecan (1:1000, Cat#28971, Cell Signaling Technology, USA), SOX-9 (1:800, Cat #82630, Cell Signaling Technology, Boston, USA), or β-actin (1:5000, Cat#sc-69879, SCBT, USA) for 12 hours at 4° C. Then, the secondary antibody (1:2000, SCBT, USA) was introduced and cultured for 90 min, followed by exposure to ECL solution.

Techniques: Western Blot

Journal: Aging (Albany NY)

Article Title: Cabozantinib prevents AGEs-induced degradation of type 2 collagen and aggrecan in human chondrocytes

doi: 10.18632/aging.205186

Figure Lengend Snippet: Silencing SOX-9 abolished the effect of Cabozantinib on type II collagen and aggrecan levels in AGEs-treated SW1353 chondrocytes. Cells were transfected with the siRNA targeting SOX-9 (siR-SOX-9), followed by stimulated with 100 μg/ml AGEs in the presence or absence of 20 μM Cabozantinib for 24 h. ( A ) Successful knockdown of SOX-9. ( B ) The protein level of type II collagen and Aggrecan was determined by the ELISA assay (n=6, *, ** P <0.05, 0.01 vs. Control group, #, ## P <0.05, 0.01 vs. AGEs group, ※ P <0.05 vs. Cabozantinib group).

Article Snippet: After separation using a 12% sodium dodecyl sulfate (SDS)-polyacrylamide gel (PAGE), proteins were moved to the polyvinylidene fluoride (PVDF) membrane, followed by loading with primary antibodies against type II collagen (1:800, Cat#28459-1-AP, Proteintech, USA), Aggrecan (1:1000, Cat#28971, Cell Signaling Technology, USA), SOX-9 (1:800, Cat #82630, Cell Signaling Technology, Boston, USA), or β-actin (1:5000, Cat#sc-69879, SCBT, USA) for 12 hours at 4° C. Then, the secondary antibody (1:2000, SCBT, USA) was introduced and cultured for 90 min, followed by exposure to ECL solution.

Techniques: Transfection, Enzyme-linked Immunosorbent Assay