Cells were microinjected using a FemtoJet 4i microinjector with micromanipulator InjectMan 4 (Eppendorf) with 80 hPa injection pressure, 0.3 s injection time, and 10 hPa compensation pressure. Each cell was injected with 200 fl of phosphate-buffered saline containing ~25,000 molecules of recSHIP2 (SignalChem) together with fluorescence marker Dextran Alexa Fluor 647 (Thermo Fisher Scientific) and 100 molecules of pKrox24(MapErk)dTomato reporter. Cells were treated for 24 hours with FGF2 (10 ng/ml), fixed with 10% paraformaldehyde, and immediately imaged for pKrox24 transactivation, using the automatic microscope TissueFAXS i (TissueGnostics) with 20× air objective. We acquired images for red and far-red fluorescence channels showing pKrox24(MapErk)dTomato reporter and far-red counterstain marker Dextran Alexa Fluor 647. Percentages of injected cells positive for dTomato were determined by Fiji free software.

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