Figure 2
From: Intratumoral heterogeneity impacts the response to anti-neu antibody therapy
Metastatic capacity of TUBO-P2J cells in vitro and in vivo . A) Percent survival of TUBO and TUBO-P2J bearing mice. TUBO (4 × 105 /mice) and TUBO-P2J (2 × 105 /mice) cells were implanted subcutaneously on the backs of mice. Fourteen days after implantation, primary tumors were surgically removed and mouse survival was recorded. B) Picture of lungs from TUBO-P2J (2 × 104 /mice) tumor bearing mouse 30 days after implantation. C) Scratch wound assay: 1 × 106 cells of TUBO or TUBO-P2J were seeded in 6 well culture plates. The surface of the culture plates was scratched mechanically with pipette tips and cell migration was evaluated 3 days later. D) Trans-well migration assay: 5 × 104 cells of TUBO and TUBO-P2J cells were added to the top plate and incubated for 4 hours. Migrated cells were counted from 5 randomly selected fields under × 40 magnification and averages were calculated. Data shown are representative of three independent experiments (mean ± SD). E) Invasion assay: 1 × 105cells of TUBO and TUBO-P2J cells were added to matrigel coated trans-well plates and incubated for 3 days. Invaded cells were counted from 5 randomly selected fields under × 40 magnification and averages were calculated. Data shown are representative of three independent experiments (mean ± SD). F) RT-PCR for expression of MMPs. G) Zymography for MMPs H) Inhibition of invasion by MMP9 inhibitor I. Invasion assay was done with indicated doses of MMP9 inhibitor by the same procedure as (E). Data shown are representative of two independent experiments (mean ± SD). * is p < 0.05 and *** is p < 0.001 versus TUBO or DMSO.