Download A Figure S7. A. Standard curve of actin quantification using silver

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.