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. 2019 Mar 27;21(4):413–427. doi: 10.1016/j.neo.2019.03.003

Figure 5.

Figure 5

LOXL2 promotes the expression and secretion of pro-lymphangiogenic factors in fibroblasts through HIF-1α. (A) The schematic flowchart shows the co-culture system for fibroblasts and tumor cells. Then detect the cell lysis and CM of the educated fibroblast cells. (B) The mRNA levels of SDF-1α and VEGF-C in fibroblast cells educated by tumor cells (top). The protein levels of SDF-1α and VEGF-C in fibroblasts CM educated by tumor cells were detected by ELLSA (bottom). (C) The protein levels of HIF-1α, LOXL2 and α-SMA in fibroblast cells educated by tumor cells were detected by Western blotting. (D) Immunoblot analysis of HIF-1α expression in the fibroblast cells transfected with specific siRNAs. (E) The effect of siRNAs on inhibiting VEGF-C, SDF-1α mRNA levels of fibroblast cells. (F) Density of lymphatic vessels in the Matrigel plug assay. Plugs containing CM from fibroblast cells and CM from fibroblasts transfected with specific siRNAs were subcutaneously injected into BALB/c mice. After 8 days, plugs were dissected and applied to IF analysis for lymphatic vessel density (left). Quantified results were shown (right; n = 6). The data are the means ± SD of three independent experiments. *P < .05; **P < .01; ***P < .001. Scale bar = 100 μm.