REF-52 fibroblasts. Cyclic AMP Fluorosensor (FlCRhR) and fura-2 AM

REF-52 fibroblasts loaded with fura-2 AM (F1201, F1221, F1225, F14185) following microinjection of Cyclic AMP Fluorosensor (FlCRhR, C6660). A time-course series of correlated images of cAMP (top row) and Ca2+ (bottom row) was obtained using simultaneous ratio-imaging techniques. The first column shows control images. The second column displays the fibroblasts one minute after treatment with forskolin, which increases the level of cAMP accumulation but has no effect on Ca2+ concentration. The third and fourth columns show the cells 17 and 38 min after forskolin addition, respectively. Over this time period, cAMP-induced nuclear localization of the fluorescein-labeled C subunit of FlCRhR can be observed (top row, right), whereas Ca2+ levels do not change. (Proc Natl Acad Sci U S A 93, 4577 (1996)) The image was obtained with the Attofluor RatioVision Digital Fluorescence Microscopy system. Image contributed by Gary Brooker, Maria DeBernardi and Scott McDonald, Atto Bioscience.

REF-52 fibroblasts loaded with fura-2 AM (F1201, F1221, F1225, F14185) following microinjection of Cyclic AMP Fluorosensor (FlCRhR, C6660). A time-course series of correlated images of cAMP (top row) and Ca<sup>2+</sup> (bottom row) was obtained using simultaneous ratio-imaging techniques. The first column shows control images. The second column displays the fibroblasts one minute after treatment with forskolin, which increases the level of cAMP accumulation but has no effect on Ca<sup>2+</sup> concentration. The third and fourth columns show the cells 17 and 38 min after forskolin addition, respectively. Over this time period, cAMP-induced nuclear localization of the fluorescein-labeled C subunit of FlCRhR can be observed (top row, right), whereas Ca<sup>2+</sup> levels do not change. (Proc Natl Acad Sci U S A 93, 4577 (1996)) The image was obtained with the Attofluor RatioVision Digital Fluorescence Microscopy system. Image contributed by Gary Brooker, Maria DeBernardi and Scott McDonald, Atto Bioscience.

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