Neat1

Transient interference of NEAT1 adopted h‐NEAT1 Smart Silencer (Ribobio, Guangzhou, China). The stable NEAT1 knockdown cell line was achieved through transfecting cholesterol modified antisense oligonucleotides NEAT1 (Ribobio). Effective NEAT1 siRNA as well as aso sequence was presented as GGGACAGACAGGGAGAGATG. Negative controls were all offered and adopted following the manufacturer's protocols.
The plasmid particularly expresses the human KRAS open reading frame (abbreviated as ORF) without miR‐193a‐3p responsive 3′‐UTR was designed and purchased from GeneCopoeia (Germantown, MD, USA). An empty plasmid was involved in negative control.

FISH assay was carried out as previously described [18 (link)]. First of all, 4% formaldehyde was added to incubate with VSMCs for 15 min and then PBS was used to wash them. Fixed VSMCs was then treated with pepsin and ethanol. Consequently, the FISH probes NEAT1 (Ribobio) was employed for mixing the dried VSMCs for 2 min in a hybridization buffer at 80°C. After dehydration, the slides were counterstained using DAPI and confocal microscope (Leica) captured the images.

HOTAIR, ZNF667-AS1 and NEAT1 probes were designed and synthesized by RiboBio (Guangzhou, China), and the probe sequences are available upon request
The probe signals were detected with a FISH Kit (RiboBio, Guangzhou, China) according to the manufacturer's instructions. Briefly, rearrangements were analyzed on 5 μm formalin fixed and paraffin embedded (FFPE) sections. Images were captured using a fluorescence microscope (Leica, Germany). At least 50 nuclei were scored for each probe and case.