Extended Data Fig. 9: Quantification of HK1 and total KRAS in tumour cell lines.

a, Top, a standard curve for the quantification of HK1 by immunoblot was generated by titrating recombinant GST–HK1 (rHK1) and probing with a rabbit monoclonal antibody (Cell Signaling Technology, C3534). Bottom, immunoblots of the indicated amounts of lysate (5,000 cells per µl). Calculations based on these results and a molecular mass of 100 kDa indicate 200,000 and 150,000 molecules per cell for A549 and HCT-15 cells, respectively. b, Top, a standard curve for the quantification of total KRAS by immunoblot was generated by titrating recombinant KRAS4B truncated at amino acid 171 and probing with a mouse monoclonal antibody (Sigma-Aldrich, WH0003845M1). Bottom, immunoblots of the indicated amounts of lysate (5,000 cells per µl). Calculations based on these results and a molecular mass of 21 kDa indicate 700,000 and 200,000 molecules per cell for A549 and HCT-15 cells, respectively. The standard curves in a, b were plotted by linear regression using GraphPad Prism v.8.1.1 and the goodness of fit is given as R².