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Fresh bovine pituitary extract (Gibco, 13028-014, 22.7 ng ml 1 ) and 50 ng ml 1 animal-free recombinant human EGF (Thermo Fisher Scientific, AF-100-15-1MG) were added to new medium when cells were split or plated

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The samples were transferred quantitatively onto spin columns (BioEcho) and washed as follows: 0.4% SDS in PBS (3 700 l), 6 M urea in ddH 2 O (2 700 l) and PBS (3 700 l) and transferred into new 1.5-ml LoBind tubes (Eppendorf) with 2 100 l X buffer (7 M urea, 2 M thiourea in 20 mM HEPES, pH of 7.5)

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