Figure 2. LEENE knockdown inhibits angiogenesis in vitro.

(A and B) HUVECs were transfected with scramble or LEENE LNA gapmers and then subjected to pulsatile flow (12 ± 1 dyne/cm²) for 24 hours. Total RNA from 3 biological replicates was subjected to RNA-seq. Significantly enriched pathways related to angiogenesis in the DEGs (scramble vs. LEENE LNA) were plotted with P values and numbers (gene counts) and percentage of genes involved (gene ratio). (B) Heatmap showing expression of select DEGs involved in pathways shown in A. LEENE, NOS3, KDR, and PGF are colored in green and red for distinction. (C–J) HMVECs were transfected with respective LNAs for 48 hours and then used for tube formation (C and D), scratch wound (E and F), and 3D spheroid-sprouting assays (G–J). Scale bars: 0.5 mm (C), 1 mm (E), and 100 μm (G). Data are represented as mean ± SEM from 3 independent experiments. *P = 0.02 based on 2-tailed Student’s t test.