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. Author manuscript; available in PMC: 2019 Jun 17.
Published in final edited form as: Oncogene. 2018 Nov 19;38(12):2123–2134. doi: 10.1038/s41388-018-0565-9

Figure 5.

Figure 5.

EGFL6 enhances cell growth and reduces apoptosis of breast cancer cells. a, Colony formation assay in T47D/shEGFL6 cultured with recombinant EGFL6 protein. Representative images and quantitative results in a bar graph are shown. b and c, 3D cell culture for sphere formation in a soft agar with matrigel was performed with T47D and T47D/shEGFL6 or MCF-7/EGFL6 cells. Representative images and quantitative results are shown. Scale bar, 50 μm. d, Detection of pAKT and pERK by Western blotting using cell lysates prepared with T47D vs. T47D/shEGFL6 and MCF-7 vs. MCF-7/EGFL6. e. EGFL6 expression in MDA-MB-231 (+EGFL6) and MCF-7 (+EGFL6) cells increased cell growth in comparison with the counterpart parental cells. Cancer cells were seeded in 96-well plates, 2000 cells/well and cell growth was measured by alamarBlue assay (Invitrogen) after 72 hours culture. f, Flow scattering plots show percentage of apoptosis cells using annexin-V assay kit in MCF-7 and MCF-7/EGFL6 cancer cells. g, Reduced cell growth in EGFL6 knockdown cells in comparison with parental T47D cells using alamarBlue assay. h, Apoptosis assay of T47D and T47D/EGFL6 knockdown cells using annexin-V by flow cytometry assay. All assays were repeated three times (n=3), * p < 0.05. Error bar, SD.