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. 2019 May 7;294(24):9631–9641. doi: 10.1074/jbc.RA119.008463

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© 2019 George et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 5. — Disruption of the NLS on SRSF1 induces cytoplasmic CLK1 localization. A, Ser-to-Ala mutations in the NLS of GFP-SRSF1. B, GFP-SRSF1^mut and CLK1-RFP co-localize to the cytoplasm using confocal microscopy. C, co-immunoprecipitation (IP) assays show that CLK1-FLAG interacts with both GFP-SRSF1 and GFP-SRSF1^mut in HeLa cell lysates. CLK1-FLAG is immunoprecipitated with an agarose-conjugated anti-FLAG antibody, and IgG represents the agarose resin control lacking the conjugated antibody. D, overexpression of GFP-SRSF1^mut induces changes in subcellular distribution of endogenous CLK1 in fractionation experiments. ImageJ was used to quantitate the amount of endogenous CLK1 in the nucleus (% Nuc) upon the expression of GFP-SRSF1 and GFP-SRSF1^mut. Scale bars, 20 μm. Cyto, cytoplasmic; Nuc, nuclear.