Sp600125 s5567

SET2 cells were kindly provided by Prof. Dr. Fabíola Attié de Castro (School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil). HEL was obtained from ATCC (Philadelphia, PA, USA). SET2 and HEL cells harboring JAK2^V617F mutation were tested and authenticated by Short Tandem Repeat (STR) matching analysis using the PowerPlex® 16 HS system (Promega, Madison, WI, USA) and the ABI 3500 Sequence Detector System (Life Technologies, Foster City, CA, USA). Cell culture conditions were performed in accordance with the recommendations of ATCC and DSMZ. All cell lines were mycoplasma-free. Ruxolitinib was obtained from InvivoGen (San Diego, CA, USA). Reversine was obtained from TargetMol (Target Molecule Corp., Boston, MA, USA). Aurora-A Inhibitor I (SML0882), AZD1152-HQPA (SML0268), NMS-P715 (475949) and SP600125 (S5567) were obtained from Sigma-Aldrich (St. Louis, MO, USA).

The S1PR1 specific inhibitor W146 (sc 296700) was purchased from Santa Cruz Biotechnology (CA, USA). The drug-dissolving agent dimethyl sulfoxide (DMSO), S1PR1 specific agonist SEW2871 (S3944), ERK/MAPK-specific inhibitor PD98059 (P215), p38/MAPK specific inhibitor SB202190 (S7067), and JNK/MAPK-specific inhibitor SP600125 (S5567) were purchased from Sigma–Aldrich (St. Louis, MO, USA). Rabbit anti-S1PR1 (EDG-1) antibody (BS2593) was purchased from Bioworld (MN, USA). Rabbit anti-chicken NLRP3 polyclonal antibody was gifted by Zhangyong Ning (South China Agricultural University). Rabbit anti-GAPDH antibody (ab181602) was purchased from Abcam (Cambridge, UK). Rabbit anti-phospho-p44/42 MAPK (Erk1/2) antibody (#9101), rabbit anti-p44/42 MAPK (Erk1/2) antibody (#4695), rabbit anti-phospho-p38/MAPK antibody (#4511), and rabbit anti-p38/MAPK antibody (#8690) were purchased from Cell Signaling Technology (MA, USA). Rabbit anti-phospho-JNK1/MAPK antibody (bs-17591R) was purchased from Bioss Biotechnology Co., Ltd. (Beijing, China). Rabbit anti-JNK1/MAPK antibodies (ab199380) were purchased from Abcam. Rabbit anti-NDV-NP polyclonal antibodies were prepared and stored in our laboratory.

Anti-LRIG2 (ab121472) was purchased from Abcam (Cambridge, MA, USA). Anti-caspase 3 (9662), anti-caspase 8 (9746), anti-caspase 9 (9502), anti-AKT (9272), anti-pS473-AKT (4058), anti-pY1068-EGFR (3777), and anti-pY1086-EGFR (2220) were purchased from Cell Signaling (Danvers, MA, USA). Antibodies against cytochrome c (sc-7159), EGFR (sc-03), MCL-1(sc-819), BCL-xL (sc-8392), BCL2A1, BCL-2 (sc-65392), BCL-W (sc-6418), BAK (sc-832), BAX (sc-493), BAD (sc-8044), BIM (sc-11425), p21 (sc-397), and β-actin (sc-47778) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-pY1101-EGFR (MAB1382) was purchased from Abnova (Taipei, Taiwan). Anti-COX IX (A21347) was purchased from Invitrogen (Carlsbad, CA, USA). LY294002 (L9908) and SP600125 (S5567) were purchased from Sigma. U0126 (662005) was obtained from Calbiochem (San Diego, CA, USA). Gefitinib (G4408) was purchased from LC laboratories (Woburn, MA, USA). Other reagents were purchased from Sigma-Aldrich unless otherwise specified.