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Figure 4 | Cancer Cell International

Figure 4

From: Soluble ephrin a1 is necessary for the growth of HeLa and SK-BR3 cells

Figure 4

Transfection of HeLa cells with full length EFNA1 (A1) or a truncated EFNA1 lacking the C-terminal signal for GPI anchor attachment (A1-ve GPI). Shown is an anti-EFNA1 western blot of lysates and conditioned media. Truncated EFNA1 accumulates in the conditioned medium whereas the full-length EFNA1 does not. B) Soluble EFNA1 in conditioned media rescues the growth of EFNA1 knockdown cells. HeLa cells stably transfected with either vector control or shRNA against EFNA1 were grown in conditioned media from either vector control or knockdown cells (own CM) or conditioned media from cells in which truncated soluble EFNA1 is overexpressed (A1-veGPI CM). C) Soluble EFNA1 in conditioned media partially rescues the growth of EFNA1 knockdown cells in semi-solid media. HeLa cells stably transfected with either vector control or shRNA against EFNA1 were grown in semi solid media that was supplemented with either conditioned media from either vector control or knockdown cells (own CM) or conditioned media from cells in which truncated EFNA1 is overexpressed (A1-veGPI CM). D) EFNA1-Fc rescues the growth of EFNA1 knockdown cells in semi-solid media. HeLa cells stably transfected with either vector control or shRNA against EFNA1 were grown in semi media that was supplemented with either EFNA1-Fc (A1Fc) or Fc alone. E) Overexpression of soluble EFNA1 (EFNA1-GPI) but not full length EFNA1 promotes the growth of HeLa cells. HeLa cells were stably transfected with pcDNA3, EFNA1 full length, or truncated EFNA1-GPI. Pools of stable clones were then counted, seeded at low density, and growth was monitored by crystal violet staining. F) Overexpression of EFNA1 inhibits the growth of HeLa cells in semi-solid media. The stable pools of HeLa cells described above were plated at the same density in semi-solid media and the number of colonies determined after two weeks of growth.

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