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Fig. 4 | Molecular Cancer

Fig. 4

From: N6-methyladenosine induced miR-143-3p promotes the brain metastasis of lung cancer via regulation of VASH1

Fig. 4

VASH1 mediates miR-143-3p induced angiogenesis via destabilization of VEGFA. a A549 or H1975 cells were transfected with vector control or VASH1 construct for 24 h, the expression of VEGFA was measured by western blot analysis (left) and quantitatively analyzed (right). b A549 control or miR-143-3p stable cells were transfected with vector control or VASH1 construct for 24 h, the expression of VEGFA was measure by western blot analysis (left) and quantitatively analyzed (right). c Tube formation was evaluated in HUVECs cultured for 12 h in medium collected from A549 cells transfected with vector or pcDNA/VASH1 and supplemented with or without 10 ng/ml rVEGFA. d A549 cells were transfected with control, miR-143-3p, vector control or VASH1 construct for 24 h, the mRNA expression of VEGFA was measured by qRT-PCR. e Analysis of VEGFA mRNA in non-ribosome portion (<40S), 40S, 60S, 80S, and polysome in A549 cells transfected with vector control or VASH1 constructs for 24 h. f A549 cells transfected with vector control or VASH1 constructs for 24 h were further treated with 10 μg/ml CHX for the indicated time periods. The expression of VEGFA was detected by western blot analysis (left) and quantitatively analyzed (right). g A549 cells transfected with vector control or VASH1 constructs for 24 h were further treated with 10 μg/ml CHX or 10 μM MG-132 for 6 h, the expression of VEGFA was detected by western blot analysis (left) and quantitatively analyzed (right). h A549 cells transfected with vector control or VASH1 constructs for 24 h in the presence of MG-132. After VEGFA was immunoprecipitated, the ubiquitination of VEGFA was examined by western blot analysis. An equal amount of VEGFA was loaded after IP according to a pre-Western blot. Data are presented as the mean ± SD from three independent experiments. *p < 0.05, ** p < 0.01 compared with control, NS, no significant. Scale bar = 100 μm

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