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. Author manuscript; available in PMC: 2014 Apr 11.
Published in final edited form as: Nat Commun. 2013;4:2956. doi: 10.1038/ncomms3956

Figure 5. FOXG1 silencing decreases BTIC-initiated brain tumour growth.

Figure 5

(a–e) Comparison of brain tumour growth in NOD-SCID mice euthanized 10 weeks after implantation of BTIC line BT048 transduced with lentivirus encoding GFP together with either non-silencing shRNA or FOXG1 shRNA #1. (a) First column shows dorsal view of a pair of representative brains. All other columns show whole-mount GFP expression in five separate implanted brains. Scale bar: 2 mm. (b) Quantification of GFP expression across the dorsal brain of implanted mice as an area of green pixels using Adobe Photoshop® (mean±SEM; P=1.43×10−4; n=5; t-test). (c) GFP expression in coronal sections through the forebrain of implanted mice. Arrows point to location of tumour cells. Scale bar: 2 mm. (d) Representation of coronal sections through different levels of the forebrain used for cell counting studies; arrows point to the corpus callosum. (e) Graph depicting the number of GFP-expressing tumour cells in the corpus callosum of the forebrain of implanted mice. Cell counts are based on six coronal sections through equivalent locations of separate brains (mean±SEM; P=1.02×10−4; n=3 brains; t-test). (f) Kaplan-Meier survival curves of mice implanted with non-silenced or FOXG1-silenced (shRNA #2) BT048 cells (non-silencing shRNA, n=8 mice; FOXG1 shRNA #2, n=7 mice). Statistical analysis (P value) is shown (Mantel-Cox test). (g) Expression of GFP and human nuclear antigen in tumour cells in coronal sections through the forebrain of mice implanted with non-silenced or FOXG1-silenced (shRNA #1) BTIC line BT025. Arrows point to location of tumour cells. Scale bar: 2 mm. (h) Kaplan-Meier survival curves of mice implanted with non-silenced or FOXG1-silenced (shRNA #1) BT025 cells (non-silencing shRNA, n=10 mice; FOXG1 shRNA #1, n=9 mice). Statistical analysis (P value) is shown (Mantel-Cox test). (i) Quantification of the neurosphere-forming ability of primary cultures derived from brain cancers formed in host mice implanted with BT025 cells expressing non-silencing shRNA or FOXG1 shRNA #1 (mean±SEM; P=2.3×10−2; n=3; t-test). (j) Western blot analysis of p21Cip1 expression in neurosphere-forming cells derived from brain cancers propagated in host mice by BT025 cells expressing non-silencing shRNA or FOXG1 shRNA #1 (two examples are shown in each case). Molecular size markers are indicated in kDa.