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. 2015 Feb 19;14(8):1164–1173. doi: 10.1080/15384101.2015.1010866

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© 2015 The Author(s). Published with license by Taylor & Francis Group, LLC

© Sharon Brookes, Sladjana Gagrica, Elaine Sanij, Janice Rowe, Fiona J Gregory, Eiji Hara, and Gordon Peters

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

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Figure 3. — Comparison of cyclin-CDK-CKI interactions at the permissive and non-permissive temperatures. (A) Cell lysates were prepared from SVts8 (SV) or BJ-TERT-tsLT (BJ) cells grown at 34°C or 6 d after shifting to 39°C. Equal amounts of protein were either analyzed directly by SDS-PAGE and immunoblotting (left panel) or after immunoprecipitation with an antibody against CDK4 (right panel). (B) Equal amounts of cell lysate (300 μg of total protein) from SVts8 cells grown at 34°C and 39°C were immunoprecipitated with antisera against CCND1, CDK4, p21^CIP1 and p16^INK4a as indicated. After fractionation by SDS-PAGE, the precipitated proteins were immunoblotted for CCND1, CDK4, p21^CIP1 and p16^INK4a as indicated. The total amounts of CCND1, CDK4, p21^CIP1 and p16^INK4a were monitored by direct immunoblotting of 30 μg of each protein sample (Total). (C) Similar analysis of CCND1 immunoprecipitates from SVts8 cells transduced with an shRNA against p21^CIP1 or non-specific shRNA.