Complete medium

Indicated ESCC cells were transfected with indicated constructs for 24 h, and then subjected to vascular mimicry formation assay. 300 µL matrigel was added into each well of the 24‐well plate. Then, 1 × 10⁵ cells in 300 µL complete medium (Lonza, Walkersville, MD, USA) were added to each well and incubated at 37 °C, 5% CO₂. After 12 h of incubation and photographs were taken.

Purchased human microvascular endothelial cells (hMVECs) were cultured in complete medium (Lonza, Basel, Switzerland). The hMVECs were used at passages 7 and 8, with a seeding density of 2 × 10⁶ cells/ml. Neurons were isolated from the brain cortex of E15 ICR mouse euthanized by CO₂ inhalation, according to procedures^{41 ,42} , and were then immediately seeded and cultured. All animal experiments were conducted in accordance with institutional guidelines and protocols approved by Korea University's Institutional Animal Care and Use Committee (KUIACUC-2017-71 and KUIACUC-2022-0028). The study was reported in accordance with ARRIVE guidelines.