Figure 3
MCF-7 cell growth in soft agar and extracellular lactate after GIT1 or MAP3K4 shRNA knockdown. The MCF-7 derived cell lines scrambled shRNA, GIT1 knockdown and MAP3K4 knockdown were cultured in a 6 well plate in soft agar for 2 weeks at 5% CO2 and 37 °C. A total of 6 samples per cell line were stained with crystal violet and pictures were taken in the ChemiDoc XRS+ System using the Coomasie Blue filter setting. Two representative pictures of each cell line are presented (Panel A). Colonies were counted with the ImageJ software using the colony count add-on. Statistical significance was assessed with non-paired Student’s t-test (Panel B). The MCF-7 derived cell lines scrambled shRNA, GIT1 knockdown and MAP3K4 knockdown were cultured in a 96 well plate and serum starved for 5 hrs then treated with CHC inhibitor for 1 hr. Then, the cells were stimulated with HRG for 16 hrs at 5% CO2 and 37 °C. After incubation, lactate measurements were taken. The graph shows the extracellular lactate concentrations after 16 hrs of HRG from two samples of different experiments. (Panel C). ***P ≤ 0.0005.
