Recombinant rat EGF was obtained from PeproTech (Cat. #400–25) and recombinant rat beta-NGF from R&D Systems (#556-NG-100). Recombinant human NRG1α (Cat. #296-HR) and NRG1β (Cat. #396-HB/CF) were obtained from R&D Systems. All ligands were reconstituted in sterile PBS containing 1 mg/ml bovine serum albumin (BSA) (Roche #03116956001). The following antibodies were used: phospho-p44/42 MAPK (ERK1/2) (pThr202/pTyr204) (E10) (Cell Signaling Technology (CST) #9106), p44/42 MAPK (ERK1/2) (L34F12) (CST #4696), Grb2 (C-23) (Santa Cruz #sc-255) and Grb2 (CST #3972), phospho-EGFR (pTyr1173) (53A5) (CST #4407), phospho-EGFR (pTyr845) (CST #2231), phospho-EGFR (pTyr1086) (CST #2220), phospho-EGFR (pTyr845) (CST #2234), hEGFR (Lab Vision Ab-15: ThermoFisher Sci MS-665-P0), EGFR (CST #2232), phospho-TrkA (pTyr674/675) (C50F3) (CST #4621), phospho-TrkA (pTyr490) (CST #9141), phospho-TrkA (pTyr785) (C67C8) (CST #4168), phospho-TrkA (pTyr785) (R&D #AF5479), TrkA (R&D #AF1056), Rap1A/Rap1B (26B4) (CST #2399), FRS2 (Fisher Sci #PA124685), phospho-ErbB4 (pTyr1242) (4C6) (Santa Cruz #sc81491), phospho-ErbB4 (pTyr1284) (21A9) (CST #4757) and anti-FLAG (CST #2044). The following kinase inhibitors were used: Gö7874 (Calbiochem #365252), erlotinib (CST #5083), and the dynamin GTPase inhibitor Dyngo 4a (Abcam #ab120689).
Kiyatkin A., van Alderwerelt van Rosenburgh I.K., Klein D.E, & Lemmon M.A. (2020). Kinetics of receptor tyrosine kinase activation define ERK signaling dynamics. Science signaling, 13(645), eaaz5267.
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