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Figure EV3. PAWS1 regulates the CK1α protein but not mRNA levels in cells PAWS1WT and PAWS1−/− U2OS cells were treated with 10 μM of the proteasome inhibitor Bortezomib for 6 h and extracts were analysed by Western blotting with the indicated antibodies. PAWS1WT and PAWS1−/− U2OS cells were treated with 50 μM Bafilomycin A1 (BafA1) for 2 or 16 h, and extracts were analysed by Western blotting with the indicated antibodies. Correlation plots of PAWS1 and CK1α protein expression from Fig 6E based on densitometry (using ImageJ) quantification of immunoblots normalised to GAPDH levels (a.u.: arbitrary units). Each dot represents a cell line. Pearson r coefficient and one‐tailed P‐values were calculated with Prism6 software. As in (C), except that PAWS1 and CK1ε protein levels were quantified and plotted. CK1α mRNA in PAWS1−/− cells rescued with PAWS1WT or GFP control analysed by quantitative RT–PCR. Data are represented as fold induction over controls and normalised internally to β‐actin control. Error bars represent ± SEM (n.s.: no statistical significance; n = 3). Relative expression of PAWS1 and CK1α transcripts in the indicated cancer cell lines was measured by RT–PCR (n = 7; error bars represent ± SEM). Correlation plot of relative PAWS1 and CK1α mRNA expression (normalised to β‐actin) in a subset of the indicated cancer cell lines. Pearson r coefficient and one‐tailed P‐values were calculated with Prism6 software. Image published in: Bozatzi P et al. (2018) © 2018 The Authors. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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