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Fig. 5. The cAMP-PKA pathway mediates the effect of glucose on Cx36 levels. INS-1 cells were incubated for 24 hours at 2 mM glucose with or without 20 µM forskolin or at 20 mM glucose with or without 10 µM H89. (A) Western-blot analysis showed that forskolin had a similar effect to glucose on Cx36 expression, whereas H89 fully blunted the effects of glucose and forskolin. Data are means ± s.e.m. of densitometric analyses of six independent experiments made in duplicate. ^**, P<0.01, ^***, P<0.001. (left) Immunolabeling of Cx36 (Cx36 recognized by fluorescein-conjugated secondary antibodies) resulted in a punctate membrane labeling in INS-1 cells (white tab represents 50 µm). (a) 2 mM glucose; (b) 20 mM glucose; (c) 2 mM glucose, 20 µM forskolin; (d) 20 mM glucose, 10 µM H89. Pictures are representative of four independent experiments. (right) Quantitations of the fluorescent dots were performed under the microscope on many images by counting the immunofluorescent dots in a square containing 100 cells. Data are means ± s.e.m. of four independent experiments. ^***, P<0.001.

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