Phospho trkb tyr816

The primary antibodies against phospho-Akt (Ser473), pan-Akt, Phospo-CREB (Ser133), pan-CREB, phospho-TRKA (Tyr 674-675)/TRKB(Tyr706-707), pan-TRKB, phospho 4EBP1 (Thr37/Thr46), pan 4EBP1, phospho FOXO1 (Ser256), pan FOXO1, phospho P70S6K (Thr389), pan P70S6K, and Caspase 3, were obtained from Cell Signaling Technology (Beverly, MA, USA). BDNF was from Santa Cruz Biotechnology (Dallas, TX, USA). phospho-TRKB (Tyr816) was purchased from Millipore (Burlington, MA, USA). The β-actin antibody was from Bioss Antibodies Inc. (Woburn, MA, USA). The appropriate horseradish peroxidase-conjugated secondary antibodies were purchased from Abcam (Cambridge, UK). For Q-PCR, Power SYBR Green PCR Master Mix and 96-well optical reaction plates were from Applied Biosystems (Thermo Fisher Scientific, Waltham, MA, USA). The primers were synthesized and sequenced by Eurofins (Milan, Italy). Beta Hydroxy Butyrate was purchased from Sigma Aldrich (Merck kGaA, Darmstadt, Germany). All standards for mass spectrometry were purchased from Sigma-Aldrich S.r.l. (Milan, Italy). All the solutions and solvents were of the highest available purity and were suitable for LC–MS analysis and purchased from J. T. Baker (Phillipsburg, NJ, USA).

Primary antibodies used in this study are as follows: antibodies against RBM4 (1:2000 for immunoblotting, 1:150 for IHC; ProteinTech, 11614-1-AP), Pax6 (1:150; Millipore, AB2237), BrdU (1:50; ABclonal, A1482), Ki67 (1:300; Abcam, ab15580), calbindin (1:200; Abcam, ab108404), NeuN (1:1000; Abcam, ab177487), cleaved caspase-3 (1:100; 5A1E, Cell Signaling Technology, 9664), BDNF (Abcam, ab108319), TrkB (1:1000; Cell Signaling Technology, 4603), phospho-TrkB Tyr816 (1:200; Millipore, ABN1381), phospho-TrkB Tyr817 (1:1500; Invitrogen, MA5-32207), FLAG® (1:2500; Sigma-Aldrich, F1804), GAPDH (1:3000; ProteinTech, 60004-1-Ig), β-actin (1:5000; ProteinTech, 66009-1-Ig) and α-Tubulin (1:3000; Millipore, 05-829). These antibodies were quality-checked by previous publications.