Fig 2: LncAKHE interacts with YEATS4.a Biotin labeled lncAKHE and anti-sense controls were used for RNA pulldown assays. Then SDS-PAGE, silver staining and mass spectrometry were conducted sequentially. YEATS4 was identified as a potential interactive protein of lncAKHE. b Biotin labeled lncAKHE and control sequences were added into sample lysates and RNA pulldown assays were conducted. c Anti-YEATS4 was added into sample lysates and RNA IP assay was performed. d LncAKHE was co-localized with YEATS4 in the nucleus of sample cells. Green lncAKHE, Red YEATS4, Blue DAPI. Scale bar = 10 µm. e Domain mapping showed that region 1201 ~ 1600 in lncAKHE was essential for its association with YEATS4. Data is from three independent experiments and expressed as mean ± SD. **p < 0.01

Fig 3: LncAKHE recruits YEATS4 to initiate NOTCH2 expression and activate NOTCH2 signaling.a LncAKHE enriched on NOTCH2 promoter by CHIRP assay. LncAKHE probes were biotin-labeled. Genomes from HCC sample lysates were sonicated into fragments of ~ 500 bp and incubated with lncAKHE probes. b YEATS4 was enriched on NOTCH2 promoter as shown by ChIP assays. c LncAKHE knockdown impaired the enrichment of YEATS4 on NOTCH2 promoter. d Overexpressing YEATS4 or lncAKHE promoted the enrichment of RNA Pol II on NOTCH2 promoter. e Overexpressing YEATS4 or lncAKHE promoted the expression of NOTCH2. f LncAKHE overexpression promoted cell proliferation and migration while inhibition of NOTCH signaling by addition of IMR-1 reversed it. Data is from three independent experiments and expressed as mean ± SD. **p < 0.01