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. 2003 Jan 6;160(1):101–112. doi: 10.1083/jcb.200209127

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Copyright © 2003, The Rockefeller University Press

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Figure 3. — Confirmation of micro-FRET detection of AKAP79 binding to CaN and PKA by YFP acceptor photobleaching imaging of FRET CFP donor quenching in fixed COS7 cells. (A) Membrane colocalization of AKAP79–CFP (blue) and CaNA–YFP (green) with micro-FRET^C (pseudo-color/gated to YFP [blue underlay]), and relief of FRET CFP donor quenching by YFP acceptor photobleaching (ΔCFP pseudo-color/gated to CFP_post [blue underlay]) observed for AKAP79WT (top) but not ΔCaN (Δ315–360) (bottom). B) Membrane colocalization of AKAP79–CFP (blue) and PKA-RII–YFP (green) with micro-FRET^C (pseudo-color/gated to YFP [blue underlay]) and relief of FRET CFP donor quenching by YFP photobleaching (ΔCFP pseudo-color/gated to CFP_post [blue underlay]) observed for AKAP79WT (top) but not ΔPKA (1–361) (bottom). Bars, ∼15 μm.