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. 2018 Aug 16;46(18):9484–9495. doi: 10.1093/nar/gky719

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© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 5. — SPOP promotes transcriptional expression of DNA repair and replication factors and is required for proper activation of CHK1 in response to replication stress. (A) U2OS, C4-2b and PC3 cells were transfected with siRNA. At 48 h after transfection, mRNA was isolated, reverse-transcribed and analyzed by qPCR with the indicated primers. (B) C4-2b, PC3, LnCaP, 22rv1 and U2OS cells were transfected with siRNA and lysed 48 h post transfection. Whole cell extracts were analyzed by western blotting with the indicated antibodies (Note: P-CHK1 blots were exposed for a prolonged time to analyse P-CHK1 in unchallenged cells). (C) U2OS, C4-2b and PC3 cells were transfected with siRNA and treated with HU for the indicated times. Whole cell extracts were analyzed by western blotting with the indicated antibodies. Error bars represent SD, n = 3. Significance was determined by a two-tailed t test: *P < 0.05; **P< 0.01.