Figure 3

HNE-modified Col II modulates adhesion molecules expression and focal adhesion kinase in human OA chondrocytes. Cells were incubated as described in Legend 1 and protein (a) and mRNA (b) levels of ICAM-1 and α1β1 integrin were analyzed in cellular extracts by Western blot and real-time RT-PCR, respectively. (b) α1β1 integrin antibody coimmunoprecipitates pFAK. One hundred micrograms of total protein extracts of chondrocytes were immunoprecipitated with mouse anti-α1β1 integrin antibody. α1β1 immunoprecipitates were immunoblotted with pFAK or α1β1 integrin antibody. (c) Protein expression of ICAM-1 was qualitatively evaluated by flow cytometry using mouse anti-human ICAM-1 and Alexa FluoR 488 goat anti-mouse IgG. Mouse isotype IgG1 was used as a negative control. (b1) untreated Col II, (b2) HNE-treated Col II at a 1:20 MR, (b3) HNE-treated Col II at a 1:200 MR. ANOVA tests and Bonferroni's multiple comparison were performed to compare each condition. The data are means ± SEM of n = 5. *P < 0.05, **P < 0.01, ***P < 0.001.