Figure 3. Morphological and molecular comparison of Pten¹¹⁷ cell lines with S2, Ras^V12 and Pten¹¹⁷; Ras^V12 cell lines.

(A–H) Cells are shown after 6 days of growth from the same starting cell number. (A) S2. Cells show the typical round morphology and are loosely attached to the surface. (B–D) Pten¹¹⁷. Pten 1 and 1A have round and spindle-shaped cells. Cells in the Pten 1 line are more loosely attached. Cells in line Pten X are primarily spindle shaped. (E–F) Ras^V12. Ras 7 and 3 have spindle shaped cells that form dense attached clusters. (G–H) Pten¹¹⁷; Ras^V12. Pten; Ras 8 and 9 have spindle shaped cells that form small clusters. (I) Western-blot analysis of cell extracts from the indicated lines. The Akt pathway is activated (pAkt) above control levels in all lines with Pten mutations. The Ras 7 line also shows elevated activation. The Erk pathway is activated (dpErk) in most lines with very low levels in S2 cells and undetectable levels in line Pten 1. Total Akt and Erk, as detected by α-Akt and α-Erk were used as loading controls [40], [41]. Akt is detected as two bands (Cell Signaling Technology) [31].