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. Author manuscript; available in PMC: 2013 Nov 1.
Published in final edited form as: Ann Neurol. 2012 Nov;72(5):766–778. doi: 10.1002/ana.23674

Figure 7. Laminin α2 is required for GSC growth.

Figure 7

For functional studies, laminin α2 shRNA were generated and validated by immunoblotting in non-GSCs, with purified laminin α2 used as a positive control (A). Growth and self-renewal of GSCs (derived from tumor 387) was compromised upon laminin α2 shRNA mediated knockdown (KD) as shown in tumorsphere formation assay (B) and ATP based growth assay (C) graphs. Laminin α2 knockdown also resulted in increased cell death as shown by Caspase 3/7 activity assay graphs (D). Addition of exogenous purified laminin α2 provided a partial rescue as shown by Caspase 3/7 activity assay graphs (E). In vivo studies where 1000 GSCs were co-transplanted with 100,000 ECs with NT or laminin α2 knock down (KD1) demonstrate that EC derived laminin α2 contributes to tumor growth as shown in Kaplan-Meier survival graph (F). *, p < 0.001 as assessed by one-way ANOVA for sphere formation assays; *, p < 0.001 for NT vs both KD1 and KD2. †, p < 0.001 for KD1 vs KD2 as assessed by one-way ANOVA for growth assays; *, p < 0.01 and **, p < 0.001 by one-way ANOVA for rescue studies; p<0.01 as assessed by long-rank analysis for in vivo studies.