Extended Data Fig. 6: VIL1 protein does not interact with PIF4 protein in vitro.
In vitro pull-down assay showing that MBP-VIL1 protein does not interact with GST-PIF4 proteins. MBP, MBP-VIL1 and MBP-SPA1 proteins were expressed in E. coli and purified using amylose-agarose beads. GST-PIF4 was also expressed in E. coli. Amylose-agarose bound MBP, MBP-VIL1 and MBP-SPA1 were used to pull-down different amount of purified GST-PIF4 protein. Pull-down samples were loaded 10% SDS-PAGE. GST-PIF4 was detected using anti-GST antibody, while the bait protein was visualized following Coomassie blue staining. The asterisks indicate each of MBP, MBP-VIL1 or MBP-SPA1 protein. The experiment was repeated at least two times independently with similar results.
