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. 2016 May 4;30(7):748–762. doi: 10.1210/me.2016-1001

Figure 7. Effects of glucose on MSH-induced CRE-dependent reporter activation are not due to direct effects on MSH signaling.

Figure 7.

A, Bradykinin-induced calcium signaling (1.0μM) was analyzed in fura 2-loaded mHypoA-2/12-CRE cells cultured with 25.0mM, 0.1mM, or no glucose for 24 hours. Data of 3 independent experiments performed in quadruplicates were compiled by setting the first value measured to 100% and expressed as the mean ± SEM. B, MSH-induced cAMP accumulation was measured in mHypoA-2/12-CRE cells cultured with 25.0mM or 0.1mM glucose for 24 hours. Data of 4 independent experiments performed in quadruplicates were compiled and expressed as the mean ± SEM. Asterisks indicate a significant difference to basal. C, Lysates of mHypoA-2/10-CRE cells, cultured for 24 hours with 0.1mM or 25mM glucose and then stimulated with 1.0μM MSH for the indicated period of time were subjected to Western blot analysis using either a phospho-specific antiserum against p-CREB or against the total histone-3 protein to control for the total protein amount. One representative blot is shown. Data of 5 independent experiments were quantified by densitometry, ratios between p-AMPK and histone signals calculated, MSH-induced CREB phosphorylation normalized to not stimulated cells, and expressed as the mean ± SEM. Hash signs indicate a significant difference to 100%.