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. 2013 Nov 20;8(11):e79798. doi: 10.1371/journal.pone.0079798

Figure 4. Expression analysis of CSC and angiogenesis markers.

Figure 4

A. Quantification of CD31 in primary tumors. Measurements are given as relative area occupied by positive signals with respect to reference area. Significant increase in CD31 is found in paclitaxel treated mice. Representative IHC images of CD31 immunostaining (200x) in tumor samples are shown. B. Treatment with paclitaxel (40 ng/mL for 72 h) increases SDF-1 and VEGF levels in LLC cells. C. ALDH1A1 mRNA levels measured by qRT-PCR in primary tumors and metastatic nodules from mice treated with the drugs. Paclitaxel increases the expression of ALDH1A1 in both locations. D. CXCR4 mRNA levels measured by qRT-PCR in primary tumors and metastasis from control and treated mice. Paclitaxel increases the expression of CXCR4 in primary tumors and metastatic nodules. Salinomycin reduces the expression of CXCR4 in primary tumors but not in metastasis. E. Boyden chamber assays using LLC cells after treatment with either vehicle, salinomycin (1 µg/mL) or paclitaxel (40 ng/mL). A very significant increase in migration is observed in cells exposed to paclitaxel, whereas a significant reduction is found in salinomycin treated cells. *p<0.05. ***p<0.001. All the in vitro experiments were repeated at least 3 independent times (each of them in triplicates).

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