JCS036467 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. The β-galactosidase liquid assay of the SAM domain. Relative values against an established positive control are shown. Nine independent colonies from CP/ILK, ΔSAM/ILK, SAM/ILK, a positive control (p22/p47) (Sumimoto et al., 1996) and a negative control (vectors alone) were subjected to β-galactosidase liquid assay as previously reported (Takeda et al., 1999). The K_D value between p22 and p47 subunit of the Nox2 complex was estimated to be 0.47 μM (Sumimoto et al., 1996). The value for SAM/ILK was not statistically significant against the negative control. *P<0.01.

• Supplemental Figure S2 -

  Fig. S2. HEK293 EphA1-GFP cells were stimulated with ephrin-A1-Fc at the indicated concentration. Whole cell lysates (WCL) were incubated with protein A Sepharose alone, washed and subjected to anti-EphA1 immunoblotting.

• Supplemental Figure S3 -

  Fig. S3. Immunofluorescent staining of EphA1-expressing cells. HEK293 Mock, WT- and KD-EphA1-FLAG cells (see Fig. 3) were fixed and stained with anti-FLAG antibody followed by detection with Cy3-conjugated anti-mouse-IgG.

• Supplemental Figure S4 -

  Fig. S4. Quantification of the spreading defect in a variety of cell types. Rat1, MCF7 and NP31 cells were transfected with GFP or WT-EphA1-GFP, subjected to spreading assays with ephrin-A1-Fc and stained by phalloidin (see Fig. 4). Spreading cells were quantified.