Skip to main content
. Author manuscript; available in PMC: 2011 May 7.
Published in final edited form as: Cell Stem Cell. 2010 May 7;6(5):421–432. doi: 10.1016/j.stem.2010.02.018

Figure 6. Integrin α6 knockdown results in a reduction in the GSC phenotype.

Figure 6

Knockdown of integrin α6 using two separate lentiviral shRNA constructs results in a decreased cell proliferation profile as assessed by the cell titer assay in T3691 (A) and T4121 (B) xenograft tumor cells. ***, p<0.001 with ANOVA comparison to non-targeting shRNA at the same timepoint; ##, p<0,01 with ANOVA comparison of shRNA1 to shRNA2 at the same timepoint. (C) Quantification of the number of tumorspheres per well and representative pictures of tumorspheres demonstrates potential self-renewal was also impaired in GSC targeted with the lentivial shRNA constructs in T4302 cells. *, p < 0.05 with ANOVA comparison to non-targeting control. (D) EdU incorporation demonstrates a decrease in proliferative capacity T4302 cells using both shRNA constructs (*** p < 0.001 with ANOVA comparison to non-targeting control, ###, < 0.001 with ANOVA comparison to shRNA1). Knockdown of integrin α6 also increases cell death as assessed by a DNA fragmentation assay (E) on T4302 xenograft tumor cells (M1 = 1 kb ladder, Un = uninfected control, NT = non-targeting shRNA control, sh1=shRNA1, sh2=shRNA2, Ctrl = staurosporine control, M2 = 100 bp ladder). Cell death was also confirmed using a caspase 3/7 assay (F) on T4121 xenograft tumor cells (***, p < 0.001 with ANOVA comparison to non-targeting control; ###, p < 0.001 with ANOVA comparison to shRNA1). (G) Kaplan-Meier survival curve demonstrate increased survival when integrin α6 is targeted with shRNA in comparison to a non-targeting control. 1000 GSCs infected with integrin α6 shRNA targeting constructs or non-targeting control were intracranially transplanted into the right frontal lobe of immunocompromised mice. *, p < 0.05; **, p < 0.01 by log-rank analysis of survival curves. (H) Representative light micrograph showing H&E staining for a control non-targeting integrin α6 1000 cell tumor showing characteristic bilateral tumor location, migrating edges, and secondary tumors, insets displayed below.