Figure 5
Lipid raft disruption through cholesterol depletion affects ERK signalling in S. mansoni somules.
Somules (~1000) were incubated in 1 or 10 mM methyl-β-cyclodextrin (MβCD) for 5–30 min and subsequently exposed to 15 ng/ml EGF. Somule protein extracts were processed for western blotting with anti-phospho PKC (Thr410) or p44/42MAPK (Thr202/Tyr204) (ERK1/2) antibodies; anti-actin antibodies were used to monitor total protein loading differences between lanes. Results are representative of those seen in two independent experiments. (b) In situ analysis of ERK activation in somules exposed to 1 or 10 mM MβCD for 20 min followed by 15 ng/ml EGF, using anti-phospho p44/42MAPK (Thr202/Tyr204) (ERK1/2) antibodies. The fluorescence intensity at the tegument (e.g. at locations ♯1 and ♯2 of a single confocal z scan) was quantified (line graph) at two separate random locations for each somule (n = 15) and mean fluorescence intensity calculated ( ± SEM; bar chart). Bar = 25 μm.
