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Download A Figure S7. A. Standard curve of actin quantification using silver
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Yap et al - Fig. S7 A B C NAC1-V5 input N3ICD-V5 input 75 kDa 75 kDa Actin in supernatant 42 kDa Figure S7. A. Standard curve of actin quantification using silver staining. Actin standards were prepared by serial dilution and separated using SDS gel electrophoresis. Silver staining was carried out and band quantification was accomplished using the BioRad QuantityOne software. Linear regression revealed that the r 2 value of the curve is 0.9767. B. NAC1 and profilin-1 (PFN1) do not demonstrate a saturable curve in a quantitative pull-down assay. C. Notch3-ICD-V5 protein demonstrated negligible binding to actin at similar molar concentrations to NAC1. This V5 protein with a similar molecular weight as NAC1 was utilized as a low affinity control to demonstrate the specificity of actin for the NAC1-V5 protein.