Skip to main content
. 2016 Oct 5;6:34504. doi: 10.1038/srep34504

Figure 8. Effects of ANGPT1 and TGFβR2 targeted silencing in cell proliferation, migration and angiogenesis.

Figure 8

(A) Western blot analysis for ANGPT1 expression in MDA-MB-231 cells transfected with scramble (lane 1), sh-ANGPT1 1.1 (lane 2) or sh-ANGPT1 1.2 (lane 3) constructs using ANGPT1 (1:1000) antibodies. Upper panel, densitometric analysis of immunodetected bands. (B) Western blot analysis for TGFβR2 in MDA-MB-231 cells transfected with scramble (lane 1), sh-TGFβR2 1.1 (lane 2) or sh-TGFβR2 1.2 (lane 3) constructs using TGFβR2 (1:1000) antibodies. Upper panel, densitometric analysis of immunodetected bands. Data were normalized using GAPDH expression. Images are representative of three independent experiments. (C) MTT assays and (D) Scratch/wound-healing assays of MDA-MB-231 cells transfected with scramble, sh-ANGPT1 1.1 or sh-TGFβR2 1.2 plasmids. (E–N) Capillary tubes development using co-cultures of HUVEC and breast cancer cells. (E) MDA-MB-231 and (F) HUVEC cells plated into matrigel with endothelial complete medium. (G) Monoculture of HUVEC in presence of VEGFA (10 ng/ml). (H) Co-culture of HUVEC with MDA-MB-231 transfected with scramble or with (I) miR-204 (30 nM). (J) Co-culture of HUVEC with MDA-MB-231 transfected cells with sh-ANGPT1 1.2, (K) sh-TGFβR2 1.1 or (L) both sh-ANGPT1 1.2 and sh-TGFβR2 1.1 plasmids. Graphical representation of (M) branch points, and (N) capillary tubes number after 24 h of co-cultures. Arrowheads indicate branch points. Arrows denote capillary-like tubes structures. Results shown are the mean of three independent experiments +/− SD. *p < 0.05, **p < 0.01 and ***p < 0.001 compared to controls. Bars represent the mean of three independent experiments ± S.D.