Development of an AC8-targeted sensor for use in MIN6.

<p>(<b>A</b>) Cartoon showing the Citrine/Cerulean sensor fused to the N-terminus of AC8 with inactivating mutations in Epac2-camps and AC8 indicated. (<b>B</b>) The cAMP sensitivity of the targeted sensor was determined <i>in </i><i>vitro</i> by addition of increasing concentrations of cAMP to crude membranes prepared from HEK293 cells expressing the sensor. EC₅₀ presented as mean±SD (n=4). (<b>C</b>) Confocal images of HEK293 and MIN6 cells expressing the global and AC8 targeted forms of the Ci/Ce sensor. (<b>D</b>) RT-PCR to identify Ca²⁺-regulated adenylyl cyclases in MIN6 cDNA and in positive control reactions.