Figure 4. Annexin II translocation is specific for saturated fatty acids.

Hs578T cells were treated as described below and DRM and DSM were prepared and resolved by SDS/PAGE and immunoblotted for annexin II as described in the Experimental section. In (A), cells were incubated for 6 h with 50 μM LCSFA (lanes 1 and 2), stearate (C_18:0) and palmitate (C_16:0) or LCUFA (lanes 3–5), oleate (C_18:1), linoleate (C_18:2) and 25 μM docosahexaenoic acid (C_22:6) as indicated. Control cells were not treated with stearate (lane 6) (representative of two separate experiments). In (B), cells were starved for 48 h and then treated with 50 μM stearate for 6 h (lane 2), 50 μM bromostearate for 6 h (lane 4). Non-stearate-treated cells were used for control (lane 1). As bromostearate was dissolved in DMSO, stearate plus DMSO is also shown for comparison (lane 3) and DMSO alone (lane 5). Immunoblots from DRM of the above stated conditions were developed for annexin II and actin as indicated (representative of three separate experiments). Densitometry results of the three experiments indicate that stearate–DMSO (lane 3) is increased versus bromostearate (lane 4) and DMSO (lane 5; P=0.0200 and P=0.0300 respectively). DMSO appears to have an effect on annexin II translocation; however, it is not statistically different from bromostearate (lanes 4 and 5) in our limited set of three experiments; P=0.2552.