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Fig. 3 | BMC Cancer

Fig. 3

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

Fig. 3

Confirming the regions of ABI1 responsible for EPS8 or SOS1 binding. Characterizing regions in SOS1 and EPS8 mediating their interaction with ABI1. a. GST-pulldown assay was performed as follows. Recombinant GST-fused beads with various ABI1 fragments were incubated with LPA-treated OVCAR3 cell lysates and subsequently analyzed by immunoblotting to detect SOS1 or EPS8 expression. Results from above experiments also identified the SH3 and poly-proline+PxxDY as the regions in ABI1 responsible for SOS1 and EPS8 interaction, respectively. b. Flag-tagged proline-rich region of SOS1 (aa:1131–1333) and Myc-tagged SH3 region of EPS8 (aa:535–586) recombinant plasmids were generated, and then co-infected with HA-tagged ABI1 into OVCAR3 cells, respectively. Cell lysates were collected after LPA stimulation. The Co-IP assay was performed to determine the regions in SOS1 and EPS8 mediating their interactions with ABI1

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BMC Cancer

ISSN: 1471-2407

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