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. 2017 Feb 13;36(26):3661–3672. doi: 10.1038/onc.2016.519

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Copyright © 2017 The Author(s)

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In GIST cells, Kit’s autophosphorylation occurs mainly on the Golgi apparatus. (a) GIST-T1, GIST882 and GIST-R8 were immuno-stained with anti-pKit^Tyr721. Phase contrast images are shown. Arrowheads indicate the perinuclear region. Bars, 20 μm. pKit^Tyr721, phosphorylation at Tyr721 in Kit. (b) GIST-T1 cells were stained with anti-pKit^Tyr721 in conjunction with the indicated antibody. Golgin97 (trans-Golgi marker), blue; GM130, (cis-Golgi marker), blue; calnexin (ER marker), blue. Lysosomes were visualized with LysoTracker Red. Insets show magnified images of the perinuclear region. Dashed lines indicate cell borders. Bars, 20 μm. (c) Pearson’s R correlation coefficients were calculated from intensity analysis of pKit^Tyr721 vs organelle markers. Results are means±s.d. (n=16–26). *P<0.05, ***P<0.001. (d) Lysates from GIST-T1 were treated with peptide N-glycosidase F (PNGase F) or endoglycosidase H (endo H) then immunoblotted. CG, complex-glycosylated form; HM, high mannose form; DG, deglycosylated form. (e) Levels of phosphorylation of the complex-glycosylated form of Kit are expressed relative to those of the high mannose form of Kit. Results are means±s.d. (n=3). **P<0.01.