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. 2009 Sep 17;284(46):31972–31981. doi: 10.1074/jbc.M109.056838

FIGURE 4.

FIGURE 4.

Effects of oxidative stress, Akt, and MEK-ERK on collagen II and Sox9 expression. Normal human chondrocytes were treated with 25 μm tBHP (added twice a day) for 2 days in DMEM/Ham's F-12 supplemented with 1% mini-ITS plus ascorbate (A, C, and E) or infected for 2 days with lentivirus expressing CA MEK or CA Akt or control vector (B, D, and F). A and B, cell lysates were collected, and the collagen II protein levels (normalized to DNA) were analyzed by enzyme-linked immunosorbent assay and presented as -fold change relative to the untreated control (Ctrl) (A) or vector (B); the data are the mean ± S.D. of three independent experiments. C–F, total RNA was isolated, and quantitative real-time PCR was performed to determine mRNA expression of collagen II (C and D) and Sox9 (E and F) using the TATA box-binding protein as a control. The mRNA expression is presented as -fold change relative to the untreated (control) or vector; the data are the mean ± S.D. of four independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.005.

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