Figure S4.

LOXL2 activates intracellular signaling pathways of LECs. (A) The protein levels of p-Akt, p-Erk, Akt, Erk in mLECs treated with LOXL2 determined by Western blotting. (B) The protein expression levels of Snail and VEGFR3 in the hLECs were detected by immunoblot. VEGF-C was used as a positive control. (C) The protein expression levels of Snail and VEGFR3 in the mLECs were detected by immunoblot. VEGF-C was used as a positive control. (D) Immunoblot analysis of Snail and VEGFR3 expression in the hLECs transfected with two specific Snail siRNAs. (E) qRT-PCR and Immunoblot analysis of Snail expression in the mLECs transfected with two specific siRNAs targeted to Snail. (F) Representative data showing the relative tube formation activity of mLECs transfected siNC and Snail-siRNAs. The data are the means ± SD of three independent experiments. Scale bar = 200 μm. (G) MCF7-LV and MCF7-LOXL2 cells were implanted into mice mammary fat pads (n = 6). Daily oral treatment with SAR131675 (30 mg/kg/d) or saline buffer started at day 8. Representative images of IF detection of lymphatic vessels (red, LYVE-1 staining) in MCF7-LV and MCF7-LOXL2 tumors (top). Quantified results of relative lymphatic vessels density were shown (bottom). (H) Representative images of immunofluorescence detection of GFP-tumor cells in lymph nodes (top). Quantified results of relative metastatic area were shown (bottom). *P < .05; **P < .01; ***P < .001. Scale bar = 100 μm.