Fig. 9
From: Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis
CXCR4 axis is activated in H460shFHC cells. a Western Blot analysis for CXCR4 expression was performed on 50 μg of total protein extract from H460shRNA and H460shFHC cells. γ-Tubulin was used as loading control. Representative data from one of three experiments. b Cells were stained with anti-CXCR4 PE (10 μL/1 × 106 cells) and evaluated by a FACS Canto II cytofluorometer. Final results represent mean ± SD of three independent experiments. Statistical significance was evaluated by Student t-test (n.s., not significant). c Western Blot analysis for pAKT and pERK expression was performed on 50 μg of total protein extract from H460shRNA and H460shFHC cells. Cells were stimulated with CXCL12 (100 ng/ml for 10 min) and treated with the AMD3100 (10 μM) one hour before their exposure to CXCL12. AKT and ERK were used as loading control. Numbers below the western blot indicates sample progression. Representative data from one of three experiments. d CXCL12 dependent-cell migration was examined in 24-well plates. Cells were placed in the upper chamber (8 μm) in the presence of AMD3100 (10 μM). Cells migrated toward CXCL12 (100 ng/ml) for 18 h. The cells were counted in ten different consecutive high power fields (magnification 200X). Final results represent mean ± SD of three independent experiments. Statistical significance was evaluated by Two-Way ANOVA (Tukey’s) (n.s., not significant, *, p < 0.05, ***, p < 0.001, ****, P < 0.0001)