Figure 2. The effect of EGFATFKDEL on RH30 and TC-71 cells in vitro.

(A and B) ³H Leucine protein synthesis assay where 1x10⁴ cells/ well were plated in triplicate and allowed to adhere overnight. RH30 (A) or TC-71 (B) cells were then pulsed for 72 hours with increasing doses (0.01-100 nM) of either EGFATFKDEL (circle) vs. BIC3 (triangle, negative control). ³H Leucine uptake was measured and data are reported as percent control response (y-axis). (C and D) shows the percent viability using Annexin V and 7AAD staining of RH30 (C) and TC-71 cells (D) treated with 2.5 μg EGFATFKDEL (open circle), compared to mitomycin positive control (triangle). Negative controls included BIC3 treated (square) and untreated cells (closed circle). Cells were harvested and analyzed at time points ranging from 12-96 hours. Above experiments are representative of 2 individual experiments.