Skip to main content
. 2011 Aug 18;1(7):869–881.

Figure 2.

Figure 2

Anchorage-dependent and -independent growth of HTK1-E6E7-HRASG12V-MYC and HTK1-K4DT-DNp53-HRASG12V-MYC cells. (A) HTK1-K4DT cells were serially infected with lentiviruses encoding DNp53 and retroviruses encoding HRASG12V, MYC or empty vectors (-). After selection, cells were harvested and subjected to SDS-PAGE. Western blots show expression of the three transgenes and suppression of p21WAF1. (B) Growth curves for DNp53-vector, DNp53-HRASG12V or DNp53-HRASG12V-MYC expressing HTK1-K4DT cells. HTK1-K4DT-DNp53-HRASG12V cells showed the fastest growth rate. Cells (2 × 104) were cultured in triplicate 12-well plates and counted every 3 days. The graphs illustrate means + s.d. (C) Anchorage independent growth of HTK1-K4DT cells expressing different transgenes. Cells (5 × 104) were seeded in 35-mm plates. After 3 weeks, colonies was counted when sized > 50 μm in diameter. The experiments were performed in triplicate and the total number of colonies in a 15 mm2 area was counted. The graphs illustrate means + s.d. Scale bars, 250 μm. (D) HTK1-E6E7 cells were serially infected with retroviruses encoding HRASG12V, MYC or empty vectors (-). After selection, cells were harvested and subjected to SDS-PAGE. Western blots show expression of the two transgenes. (E) Growth curves for vector, HRASG12V or HRASG12V-MYC expressing HTK1-E6E7 cells. HTK1-E6E7-HRASG12V cells showed the fastest growth rate. Cells were grown as described in (B). (F) Anchorage independent growth of HTK1-E6E7 cells expressing different transgenes performed as for (C). Scale bars, 250 μm.