Membrane, cytoplasmic, nuclear, and whole cell extracts were prepared as described in[
23,
97,
98]. For immunoprecipitation membrane and cytoplasmic extract fractions were re-mixed. Co-immunoprecipitation assays were performed as described previously[
23,
97,
98] using the following DTX3L and ARTD9 specific antibodies: rabbit anti-DTX3L antibody Cat.No.: D9644-01B), rabbit anti-DTX3L antibody (Bethyl Laboratories, Inc., Cat.No.: A300-833A,) and rabbit anti-ARTD9 antibody (Chemicon/EMD Millipore, Cat.No.: AB10619, Lot No.: LV1409682). All antibodies used for immunoprecipitation analysis were covalently coupled to tosyl-activated Dynabeads (Invitrogen) according to the manufacturers’ protocols. Immunoblot analysis and immunofluorescence microscopy were performed as described in[
23,
97,
98] using the following primary antibodies: Rabbit anti-DTX3L (US Biological, Cat.No.: D9644-01B), rabbit anti-DTX3L, (Bethyl Laboratories, Inc., Cat.No.: A300-833A), rabbit anti-ARTD9 (EMD Millipore, Cat.No.: AB10618), rabbit anti-ARTD9 antibody (EMD Millipore, Cat.No.: AB10619), mouse anti-ARTD2 (EMD Millipore, Cat.No.: MABE18), rabbit anti-ARTD3 (Aviva Systems Biology Corp., Cat.No.: OAAB03449), rabbit anti-ARTD10 (Aviva Systems Biology Corp., Cat.No.: ARP42810_P050), rabbit anti-ARTD12 (Aviva Systems Biology Corp., Cat.No.: OAAB03451), rabbit anti-ARTD11 (Abgent, Cat.No.: AP6297a), rabbit anti-ARTD13 (GeneTex, Cat.No.: N3C2), anti-STAT1α/β (RabMab, Epitomics, Cat.No.: 2728–1), anti-pSTAT1α/β(Y701) (RabMab, Epitomics, Cat.No.: 2825–1), anti-pSTAT1α(S727) (RabMab, Epitomics, Cat.No.: 3324–1), anti-STAT2 (RabMab, Epitomics, Cat.No.: 1513–1), anti-STAT3α (RabMab, Epitomics, Cat.No.: 3566–1), anti-pSTAT3α(S727) (RabMab, Epitomics, Cat.No.: 2236–1), anti-STAT5 (RabMab, Epitomics, Cat.No.: 1289–1), anti-pSTAT5(S726) (RabMab, Epitomics, Cat.No.: 5734–1), anti-STAT6 (RabMab, Epitomics, Cat.No.: 1505–1), anti-PTPN1 (RabMab, Epitomics, Cat.No.: 3774–1), anti-PTPN2 (RabMab, Epitomics, Cat.No.: 5790–1), anti-pJAK1 (RabMab, Epitomics, Cat.No.: 6518–1), anti-JAK1 (RabMab, Epitomics, Cat.No.: 2856–1), anti-IFNGR1 (RabMab, Epitomics, Cat.No.: 5697–1), anti-IFNGR2 (RabMab, Epitomics, Cat.No.: 7932–1), anti-IRF1 (RabMab, Cell Signaling Technology, Cat.No.: 8478), anti-STAT3α/β (RabMab, Cell Signaling Technology, Cat.No.: 12640), rabbit anti-pSTAT2(Y690) (St. Cruz Biotechnology, Inc., Cat.No.: sc-21689-R), rabbit anti-pSTAT6(Y641) (St. Cruz Biotechnology, Inc., Cat.No.: sc-101808) and mouse anti-tubulin (SIGMA, Cat.No.: T5618). The rabbit anti-ARTD8 antibody was a generous gift from Dr. Avraham Raz (Karmanos Cancer Institute, School of Medicine, Wayne State University, Detroit, Michigan 48201, USA[
99]). Immunofluorescence analysis was performed with an automated inverted research microscope system (Leica DMI6000B, Leica Microsystem). Composite images were generated by Adobe Photoshop software. Quantification of immunoblots was performed using the GelEval software (FrogDance Software Inc.) and mean value ± SE was calculated and plotted into graphs using the GraphPad Prism 5 software (GraphPad Software, Inc.).