Figure 2. Decreased GJIC and increased survival in v-Src expressing cells.

(A) Visualization of Lucifer Yellow dye transfer after scrape loading in wt and Src1 cells. (B) FACS assay for GJIC. Cells were pre-loaded with 2 fluorescent dyes: calcein, which can pass through gap junctions, and DiI, which cannot. The pre-loaded cells were then overlayed on a monolayer of unstained cells and incubated to allow time for calcein dye transfer via gap junctions. After 3 h, the mixed population was analyzed for calcein and DiI content by FACS. Cells containing both calcein and DiI represent the pre-loaded population. Cells containing only calcein (lower right quadrant) were deemed to have received the dye by GJIC. (C) Quantification of the percentage of unloaded cells that acquired calcein, indicative of gap junction transmission. (D) wt, Src1, or Src2 cells were treated at either high (30,000 cell/cm²) or low (500 cells/cm²) density, as indicated, with 2.5 μg/ml of cisplatin. One set of wt cells was also treated with lindane at 50mg/ml as indicated. For clonogenic survival of cells treated at high density, cells were treated for 1 h at high density, trypsinized and then serially diluted for the colony formation analysis.