Stimulation of proliferation and ERK1/2 activation by FBS.

<p>(a): Naïve ADSCs were plated in 24-well plates and grown to 80% confluence, after which they were cultured in DMEM + 1% FBS for 24h. Cells were subsequently treated for 24h with DMEM supplemented with FBS at the concentrations given, and proliferation was quantified by means of [³H]dT incorporation. (b): Naïve ADSCs were plated in 24-well plates and grown to 100% confluence, upon which they were treated for 7 days with vehicle control (0.1% EtOH) or with OM to generate ADSC-OBs, which were subsequently treated for 24h with OM supplemented with FBS at the concentrations indicated. Proliferation was quantified as for (a). (c,d): Western blotting for total ERK1/2 and phospho-ERK1/2 was subsequently performed on cell lysates. (c): Naïve ADSCs were plated in 100mm dishes and grown to 80% confluence, after which they were cultured in DMEM + 1% FBS for 24h. Cells were subsequently treated with DMEM + 5% FBS for the time-points indicated, and cell lysates were prepared. (d): ADSC-OBs were generated as for (a) and subsequently treated with OM + 20% FBS for the time-points indicated, after which cell lysates were prepared. Panels a and b show the representative results of three independent experiments, performed in triplicate. Panels c and d show the representative results of three independent experiments. Different lower-case letters indicate statistically significant differences (P < 0.05).