The pcDNA3.1-PKACα-YFP plasmid was a gift from Manuela Zaccolo from the University of Padua, previously described [22 (link)]. pRK5 and pRK5-Fyn constructs [23 (link)] were acquired from Addgene (Cambridge, MA, USA): human pRK5-Fyn WT (wild type), pRK5-Fyn KD (K299M, kinase dead), and pRK5-Fyn ΔSH3 (deletion of 76-141). The pcDNA3-YFP plasmid and pcDNA3.1-EV were obtained from Addgene (Cambridge, MA, USA). The pFlag-CMV2 LARP4 and pFLAG-CMV2 plasmids were a gift from Richard Maraia from the National Institute of Health, previously described [24 (link)]. The Fyn-Y3D mutant (Fyn-Y3D), possessing three tyrosine-to-aspartate mutations in the SH2 domain (Y185D, Y213D, and Y214D), was described previously [25 (link)].
The following primary antibodies were used for immunoblotting: α-Fyn (FYN3) and α-PKACα (C-20) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA), α-phosphotyrosine (4G10) was purchased from EMD Millipore (Billerica, MA, USA), α-Myomegalin was obtained from Thermo Scientific (Waltham, MA, USA), α-AKAP9 and α-CDK5RAP2 were obtained from Bethyl Laboratories (Montgomery, TX, USA), and α-tubulin was purchased from Cell Signaling Technology (Danvers, MA, USA). Secondary antibodies (α-mouse-HRP and α-rabbit-HRP) were purchased from Jackson ImmunoResearch (West Grove, PA, USA).
The following primary antibodies were used for immunoblotting: α-Fyn (FYN3) and α-PKACα (C-20) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA), α-phosphotyrosine (4G10) was purchased from EMD Millipore (Billerica, MA, USA), α-Myomegalin was obtained from Thermo Scientific (Waltham, MA, USA), α-AKAP9 and α-CDK5RAP2 were obtained from Bethyl Laboratories (Montgomery, TX, USA), and α-tubulin was purchased from Cell Signaling Technology (Danvers, MA, USA). Secondary antibodies (α-mouse-HRP and α-rabbit-HRP) were purchased from Jackson ImmunoResearch (West Grove, PA, USA).