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Figure 7


Fig. 7. (A) Western blot analysis showing the differences in short-term glucose-induced ERK1/2 activation between B1 and C3 sublines and the stimulatory effect of latrunculin B pre-treatment on the short-term phosphorylation of ERK1/2. (B) Immunofluorescence analysis of the subcellular localisation of activated phosphoERK1/2 (green) and the actin cytoskeleton (red) on B1 (top panels) and C3 cells (bottom panels) before and after 5 minutes stimulation with 20 mM glucose. Nuclei (blue) were stained with DRAQ5TM. No immunoreactivity was observed using control rabbit IgG (not shown). Bars, 10 µm. (C) PhosphoERK1/2 accumulates at the tips of actin fibres in the vicinity of the plasma membrane (white arrows) after 20 mM glucose (left) and 16.7 mM glucose (right) stimulation in B1 cells. Bar, 10 µm. (D) Inhibition of ERK1/2 activation by PD98059 has no effect on the actin cytoskeleton remodelling properties of B1 or C3 cells as shown by confocal analysis of phalloidin-labelled cell samples + PD98059 before and after 5 minutes glucose stimulation. Bars, 10 µm.





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