Figure 3
Human insulin and IGF-I stimulate PKC and ERK activation in S. mansoni somules.
(a) 72 h in vitro-cultured somules, starved overnight, were exposed to insulin (1 μM; 30, 60 min) and phosphorylated (activated) PKCs (p-PKC) and ERKs (p-ERK) detected by western blotting with anti-phospho PKC (Ser660) or (Thr410) and anti-phospho p44/42MAPK (Thr202/Tyr204) (ERK1/2) antibodies. Control somules (in BME) remained untreated. Band intensities were quantified and mean relative change in phosphorylation (n = 6, ± SEM; graph), normalised for actin levels, was calculated with respect to BME controls that were assigned a value of 1; **p ≤ 0.01, ***p ≤ 0.001. (b) In situ localization of activated PKC and ERK (green) within 72 h somules following insulin treatment using anti-phospho PKC (Ser660), anti-phospho p44/42MAPK (Thr202/Tyr204) (ERK1/2) and AlexaFluor 488 antibodies and confocal microscopy. (c) Detection of activated PKC in response to IGF-I exposure with time using anti-phospho PKC (Ser660) antibodies. Representative micrographs of somules are shown as z-axis maximum projections. Bar = 25 μm.
