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. 2015 Dec 17;7(4):4890–4902. doi: 10.18632/oncotarget.6640

Figure 3. NOTCH2 is required for PKCδ-associated mesenchymal transformation.

Figure 3

A, B. qRT-PCR for NOTCH isoforms (A) and the ligands (B) in U87 GBM cells transfected with control or PKCδ siRNAs. C, D. Immunocytochemistry for NOTCH2 (C) and ligands JAG1 and -2 (D) in U87 GBM cells transfected with control or PKCδ siRNAs. E. Western blot analysis for NOTCH2 and its ligands JAG1 and -2 in U87 GBM cells transduced with MFG or HA-PKCδ. F, G. Migration and invasion assay in U87 GBM cells transfected with NOTCH2 siRNAs (F) or treated with γ-secretase inhibitor (GSI) (G), as compared to control. H. Infiltration of U87 GBM cells transfected with control or NOTCH2 siRNAs in collagen-based matrix 3D culture system. Scale bar, 100 μm. I. Migration and invasion assay in U87 GBM cells transfected with control siRNAs or siRNAs against JAG1 or -2. J. Western blot analysis for NICD2 in U87 GBM cells transfected with control siRNAs or siRNAs against JAG1 or -2. K, L. Western blot analysis for CDH2, SNAI2 and ZEB1 (K), and immunocytochemistry for CDH2 (L) in U87 GBM cells transfected with NOTCH2 siRNAs or treated with GSI. M, N. Immunohistochemical staining (M) and qRT-PCR (N) for NOTCH-2, JAG1 and -2 in orthotopic xenograft tumors formed by U87 GBM cells transduced with control (pSuper) or PKCδ shRNAs. Scale bar, 200 μm. O, P. qRT-PCR for NOTCH-2, JAG1 and -2 in U87 GBM cells transfected by control siRNAs or siRNAs against SRC (O) or STAT3 (P). Q. Western blot analysis for NICD2 in U87 GBM cells transfected by control siRNAs or siRNAs against SRC or STAT3. β-actin was used for a loading control. *, P < 0.05 versus control; **, p<0.01 versus control.