Tgx 221

3-Methyladenine (IC50 = 4.5 mM), Wortmannin (IC50 = 3 nM) and IC-87114 (PI3K subunit p110δ, IC50 = 0.5 µM)) were purchased from Cayman Chemical (Ann Arbor, Michigan 48108, USA). LY294002 (IC50 = 1.0 µM), another PI3K inhibitor, was obtained from the Invivogen (San Diego, CA 92121, USA). The PI3-Kinase p110 α subunit inhibitor (B-0304, IC50 = 2 nM)), and γ inhibitor B-0302(IC50 = 250 nM) were from the Echelon (Salt Lake City, UT 84108, USA). The selective inhibitor of the P13K subunit p110 β, TGX-221(IC50 = 5 nM) was from EMD Millipore (Billerica, Massachusetts 01821, USA). Rabbit anti-IRF-7 (H-246) was purchased from Santa Cruz Biotechnology (Dallas, TX 75220 USA). Rabbit anti-GAPDH (Cat #5174), IRF-3 (Cat # 4302) and NF-κB p65 (Cat #4764) antibodies were available from Cell Signaling Technology (Danvers, MA 01923 USA). Goat anti-MCM2 (Cat # A300-122) was from Bethyl Laboratories (Montgomery, TX 77356, USA). The siRNA oligonucleotides were purchased from Ambion of Thermo Fisher Scientific. The sequences are p110b (ID: s93108, 5′-CAUAGAUUUUGGGCAUAUUTT-3′; ID: s93107, 5′-GCAUAGCUGGUCUUCGUUUTT-3′), p110d (ID: s71605, 5′-CGAUGAAGCUGGUUGUUCATT-3′; ID: s71606, 5′-GCCGAAAAGUGAAUGCUGATT-3′), and Vps34 (ID: 165687, 5′-GCUUGCUCGGAGCUUAAGATT-3′; ID: 165688, 5′-CGAAGCUCUAAUUUCAUGUTT-3′).

Ouabain and phalloidin were purchased from Sigma-Aldrich (St. Luis, MO, USA). The EGFR inhibitor Iressa was purchased from Tocris (Bristol, UK). Src inhibitor AZD0530, MEK inhibitor PD0325901, and p38MAPK inhibitor VX702 were purchased from Selleckchem (Houston, USA). Src inhibitor PP2 and PI3K inhibitor TGX221 were purchased from EMD Millipore (Darmstadt, Germany). Anti-Src antibody and anti-phospho Y416-Src antibody were a gift from Dr. Don Fujita at the University of Calgary. Anti-p130 antibody was purchased from Abcam (Cambridge, MA). Rabbit polyclonal anti-Na,K-ATPase and mouse monoclonal antibody 4G10 were purchased from EMD Millipore. Anti-β-actin antibody was purchased from Sigma-Aldrich. Anti-ninein antibody was characterized previously[20 (link)]. HRP-conjugated secondary antibody and Cy3- and Alex488-labeled secondary antibodies were purchased from Jackson ImmunoResearch (West Grove, PA) and Molecular Probes (Eugene, OR), respectively. Anti-ERK antibody was purchased from Cell Signaling Technologies (Danvers, MA) and Fluorescent-labeled Ouabain was purchased from ThermoFisher Scientific (Waltham, MA).

FR900359 was isolated as described previously (Schrage et al., 2015 (link)). Salts and drugs have been purchased from Sigma-Aldrich (Germany) with the exception of PTX and TGX 221, which were obtained from Merck Millipore (Calbiochem) (Germany) and PI-103 from Enzo Life Sciences. The PI3K inhibitors TGX-221, AS-252424, and PI-103 were administered at submaximal concentrations to achieve biological effects (Ali et al., 2008 (link); Tsvetkov et al., 2016a (link)). All drugs were freshly dissolved on the day of the experiment according to the material sheet. The relaxin peptides have been purchased from Sigma-Aldrich (Germany). Relaxins were dissolved in water. The following concentrations were used: relaxin-1 at 10 pM or at 100 pM, relaxin-2 at 10 pM or at 100 pM or from 1 pM to 10 nM, relaxin-3 at 10 pM or at 100 pM.