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. 2002 Feb;135(3):609–618. doi: 10.1038/sj.bjp.0704480

Figure 6.

Figure 6

IL-1β and cyclic AMP-agonists increase PDE4B2 protein in human OA chondrocytes. Chondrocytes cultured in alginate beads were exposed to vehicle, 200 pg ml−1 IL-1β in the presence or absence of 10 μM Indomethacin or 10 μM Forskolin and 1 mM IBMX, or 1 μM PGE2, or 1 μM Piclamilast for 12 h. Cellular extracts were prepared and immunoblotting was performed with rabbit polyclonal PDE4A, B or D antibodies as described in the Methods section. Each lane was loaded with 20 μg (for PDE4B and D) or 50 μg (for PDE4A) protein. (A) Antibody to PDE4A, lane 1 - recombinant human (rh) PDE4A4, lane 2 - control, lane 3 - IL-1β, lane 4 - IL-1β+Forskolin+IBMX. (B) Antibody to PDE4B, lane 1 - rh PDE4B2, lane 2 - control, lane 3 - IL-1β, lane 4 - IL-1β+Forskolin+IBMX, lane 5 - IL-1β+Indomethacin, lane 6 - PGE2. (C) Antibody to PDE4B, lane 1 - rhPDE4B2, lane 2 - control, lane 3 - IL-1β, lane 4 - IL-1β+1 μM Piclamilast. (D) Antibody to PDE4D, lane 1 - rh PDE4D3, lane 2 - control, lane 3 - IL-1β, lane 4 - IL-1β+IBMX+Forskolin, lane 5 - rhPDE4D5. The depicted blot is representative of 3 – 6 experiments.