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. Author manuscript; available in PMC: 2018 Sep 15.
Published in final edited form as: Cancer Res. 2017 Jul 7;77(18):5011–5025. doi: 10.1158/0008-5472.CAN-16-2704

Figure 2. HuR regulates PARG mRNA expression.

Figure 2

(A) mRNP-IP assay performed with cytoplasmic fraction of MIA PaCa-2 cells treated with IC50 doses of veliparib (12μM) and olaparib (9μM) for 12hr, α-Tubulin used as a loading control for the input and a negative control for the IP samples, Lamin A/C used as a control to detect nuclear contamination in the input. (B) The relative binding of PARG mRNA to HuR, normalized to respective IgG controls, as determined by RT-qPCR using 18S rRNA as a loading control, dCK as positive control and PARP-1 as negative control. (C) HuR- silenced MIA PaCa-2 cells were treated with actinomycin D (5 μg/ml) for the indicated times. PARG, GAPDH and PARP-1 mRNA stability was assayed by RT- qPCR using 18S rRNA as a loading control. (D) RT- qPCR indicating HuR and PARG mRNA expression in HuR-silenced MIA PaCa-2 cells incubated in the presence of olaparib for 24hr. (E) PARG expression in DDR- P MIA PaCa-2 and DDR- D Capan-1 and Hs 766T cells treated with veliparib for indicated time points.