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. 2017 Apr 25;116(11):1425–1435. doi: 10.1038/bjc.2017.116

Figure 3.

Figure 3

Hh pathway regulates the production of pro- and anti-angiogenic secreted factors. (A) Capillary tubes formation assay on HUVEC cells grown on Matrigel in the presence of conditioned growth media of TNBC (MDA-MB-468) cells treated with NVP-LDE225 (2.5 μM) or bevacizumab (1 μM).Cells were observed under an inverted microscope, and pictures were taken at T0 hours and after 3 h. Scale bars, 100 μm. (B) Quantitative analysis of VEGF-A in conditioned media (CM) of MDA-MB-231, SUM-159, SUM-149, HCC70 and MDA-MB-468 cells treated with NVP-LDE225 (2.5 μM) or bevacizumab (1 μM). (C, D) Quantitative analysis of (C) sVEGFR2 and (D) THBS1 in CM of MDA-MB-231, SUM-159, SUM-149, HCC70 and MDA-MB-468 cells treated with NVP-LDE225 (2.5 μM), as performed by using a multiplexed immunoassay. (E, F) Relative expression of (E) sVEGFR2 and (F) THBS1 mRNA in MDA-MB-231, SUM-159, SUM-149, HCC70 and MDA-MB-468 treated with NVP-LDE225 (2.5 μM), as performed by real-time RT–PCR (qRT-PCR) analysis. Data were calculated with mean cycle threshold (CT) values, normalised to endogenous control. Data represent the mean (±s.d.) of three independent experiments, each performed in triplicate. Bars, s.d. Asterisks indicate statistical significance, as determined by the Student t-test (*P<0.05, **P<0.01, ***P<0.001).