Fig. 3
From: Deciphering the complex role of thrombospondin-1 in glioblastoma development
THBS1 controls invasion and growth of P3 tumour. a P3 cells were transduced with shRNA control or shRNA THBS1 (−1 and −2). P3 spheroid invasion was measured in collagen I gels after 24 h. Arrows represent the migration distance from the spheroid core. Scale: 50 µm. The graph represents the invasion results as means ± s.d. of four independent experiments each done in 6–8 replicates for each condition. ****P < 0.0001 (ANOVA). b Control shRNA or THBS1 shRNA-transduced P3 cells were xenotransplanted into Ragγ2 C−/− mice. Representative images of tumours with invasive areas are shown by Nestin staining (grey) with schematic representations (invasion in red dashed lines and tumour mass in black dashed lines). Scale: 200 µm. The graphs on the right represent the number of small (<10 µm), medium (between 10 and 20 µm), large blood vessels (>20 µm) (upper panel) and the invasion area (lower panel) of control shRNA and THBS1-1/-2 shRNA-transduced P3 tumours (average of 5 tumours with 8 sections/tumour analysed; * means comparison of small vessels; # means comparison of medium size vessels). The graphs represent results as means ± s.d (n = 5 tumours analysed per group). *P < 0.05; #P < 0.05; ##P < 0.01; ns, not significant (ANOVA). c Kaplan–Meier survival curves of xenotransplanted mice with shTHBS1-1 (orange line), shTHBS1-2 (red line) or shControl (grey line) transduced P3 cells, based on the presence of neuropathological features (n = 8 mice per group); P-values were calculated with log-rank test, **P-value < 0.01, ***P-value < 0.001. d P3 cells were transduced with pLA57-control or pLA57-THBS1 lentiviral constructs. Analysis of THBS1 and Tubulin expression in protein extracts from pLA57-control or pLA57-THBS1 P3 cells by western blot (Upper panel) and by immunostaining (lower left panel), THBS1 (green) and F-actin (red) and nuclei (blue). Scale bar: 10 µm. pLA57-control or pLA57-THBS1 P3 cell spheroid invasion was measured in collagen I gels after 24 h. The graph represents the results as means ± s.d of three independent experiments each done in 6–8 replicates for each condition, **P < 0.01 (Student t-test)
