BRAF gene duplication constitutes a mechanism of MAPK pathway activation in low-grade astrocytomas
J. Clin. Invest. Stefan Pfister, et al. 118:1739 doi:10.1172/JCI33656 [Go to this article.]

Figure 5
Stable silencing of BRAF expression in pilocytic astrocytoma cells. (A) Proliferation of NCH492 pilocytic astrocytoma cells after shRNA-mediated silencing of BRAF using 3 different shRNAs as assessed by MTT assay 24 hours after plating equal numbers of cells. Nontargeting shRNA was used as a reference. Severe growth inhibition was observed for all 3 shRNAs targeting BRAF. Error bars represent the SD between replicates. (B) Microscopic examination of the same samples at the time of MTT analysis showing growth arrest of cells upon BRAF knockdown. Original magnification, ×10. (C–F) Cell-cycle analysis 24 hours after plating equal numbers of cells either carrying nontargeting shRNA (C) or one of the shRNAs targeting BRAF (D–F). As observed for the treatment with MEK1/2 inhibitor, cells accumulated in the G2/M phase of the cell cycle.