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. 2019 Mar 11;47(9):4495–4508. doi: 10.1093/nar/gkz166

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© The Author(s) 2019. 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 License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — USP17 expression is mitogen inducible and up-regulates ELK-1 target genes. (A) HeLa cells were treated with TPA and MEK inhibitor U0126 as indicated. RNA was isolated and analysed by qRT-PCR for USP17 mRNA. Results are from four biological repeats and each performed in triplicate. (B) Lysates from HeLa cells treated with TPA for 0, 30 and 60 min were analysed by SDS-PAGE (5–20%) and immunoblotting with antibodies indicated. (C–I) HEK293T cells were transfected with increasing amounts of vector for USP17 or USP22. RNA was isolated after 48 h and analysed by qRT-PCR for CFOS (C), EGR1 (D), IER2 (E), EGR2 (F), MCL1 (G), VEGF (H) or CCND1 (I) mRNA. All mRNA levels were normalized to GAPDH and represent means from three independent experiments, each assayed in triplicate. (J) HEK293T cells were transfected with Myc.USP17 and shRNA for ELK-1 as indicated. RNA was isolated after 48 h, and CFOS mRNA was analysed by qRT-PCR. Levels are presented as fold ratio of CFOS/GAPDH with USP17 normalized to 1 and represent means from three independent experiments, each assayed in triplicate. (K) Lysates from HEK293T cells from (J) were analysed by SDS-PAGE (5–20%) and immunoblotting with antibodies indicated. Significance is defined in Materials and Methods.