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. 2015 Nov 30;4(11):e176. doi: 10.1038/oncsis.2015.38

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Copyright © 2015 Macmillan Publishers Limited

Oncogenesis is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Hck disrupts Gli–Sufu interactions. (a–c) Hck activates Gli1 downstream of Kif3a in a Sufu-dependent fashion. (a) NIH3T3 cells, (b) immortalized Kif3a^−/− MEFs and (c) Sufu⁻^/− MEFs were co-infected with a control or with lentiviruses designed to express Gli1 and/or Hck. Endogenous Gli1 levels measured using primers in the 5′-UTR were determined by RT–qPCR. Presented are means plus s.d.; n=3. Significance was determined by Student's t-test; **P<0.01. (d) Hck disrupts Gli1–Sufu interactions. NIH3T3 cells were transfected with constructs expressing HA-Gli1 and Flag-tagged Sufu in the presence or absence of exogenous HA-Hck. Cell lysates were precipitated with anti-Flag antibodies and western blotted with antibodies against HA or Flag. The western blot bands were quantified with NIH Image J software and the percentage of Gli1 precipitated with Flag-Sufu was compared with the input. (e) NIH3T3 cells were transfected with constructs expressing HA-Gli1/2 and Flag-Sufu in the presence of exogenous HA-Hck or kinase-inactive Hck-K269E. Cell lysates were precipitated with anti-Flag or anti-HA antibodies and western blotted with antibodies against HA, Flag or 4G10 antibodies. Quantifications of western blot are shown below. qPCR, quantitative PCR.