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Fig. 2 | BMC Cancer

Fig. 2

From: Inhibitory short peptides targeting EPS8/ABI1/SOS1 tri-complex suppress invasion and metastasis of ovarian cancer cells

Fig. 2

Characterizing regions in ABI1 mediating its interaction with SOS1 and EPS8. a. ABI1 was divided into the following regions: WAB (aa:1–79), SNARE (aa:54–108), HHR (aa:108–153), proline-rich (aa:153–331), poly-proline (aa:331–384) and SH3(aa:384–508). b. The plasmids containing HA-tagged various regions of ABI1 above were generated, using the pCDH-CMV-MCSEF1-Puro vector. The empty vector was used as control. c. The plasmids with different regions of ABI1 were co-infected into OVCAR3 cells with Flag-tagged SOS1 or Myc-tagged EPS8, respectively. Cell lysates were collected after LPA stimulation. Co-IP was performed to map out the regions in ABI1 responsible for ABI1-SOS1 or ABI1-EPS8 interactions. The results were quantified by ImageJ. The band density of each sample were normalized by cell lysis that detected the same protein and compared with control. d. We re-divided the C-terminal of ABI1 into the following regions: poly-proline+SH3 region (aa:331–508), poly-proline+PxxDY (aa:331–425)、poly-proline (aa:331–384)、PxxDY (aa:414–425). HA-tagged plasmids containing those regions were generated. Co-IP were performed to further identify which region mediated the ABI1-EPS8 interaction

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