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Fig. 1. Expression of FP-tagged PP1 isoforms in transiently transfected mammalian cells. (A) Cartoon depicting the plasmids constructed for expression of EGFP, EYFP or ECFP PP1 ${alpha}$ , ß/ ${delta}$ and ${gamma}$ . (B) Western blot showing the bands detected with anti-PP1 antibodies in lysates (30 µg total protein) from non-transfected HEK293 cells (lane 1), and from cells transiently expressing EYFP alone (lane 2), EYFP-PP1 ${alpha}$ (lane 3), EGFP-PP1ß/ ${delta}$ (lane 4) or EYFP-PP1 ${gamma}$ (lane 5). Arrows point to expressed and endogenous PP1. (C) Western blot showing the bands detected with anti-GFP antibodies in the same lysates. (D) In vitro phosphorylase a phosphatase activity associated with EYFP, EYFP-PP1 ${alpha}$ , EGFP-PP1ß/ ${delta}$ , EYFP-PP1 ${gamma}$ and EYFP-PP1 ${gamma}$ (H125A) immunoprecipitated from these lysates using anti-GFP/YFP antibodies.

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