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. 2018 Nov 10;8(20):5676–5689. doi: 10.7150/thno.28538

Figure 4.

Figure 4

LncRNA DANCR stabilizes HIF-1α through interacting with NF90. (A-C) Hypoxia- and HIF-1α-related biological function enriched using gene set enrichment analysis (GSEA) in SUNE-1 cells transfected with DANCR siRNA or scramble control. P < 0.05, FDR ≤ 0.25. NES: normalized enrichment score. (D-E) The protein (upper panel) and mRNA (lower panel) levels of HIF-1α in HONE-1 (D) and SUNE-1 (E) cells transfected with DANCR-siRNAs, NF45-siRNAs, NF90-siRNAs or scramble control under hypoxic conditions. (F-G) Reduction of HIF-1α mRNA stability in DANCR knockdown HONE-1 (F) and SUNE-1 (G) cells as compared to control cells. Cells were treated with 1 μg/mL actinomycin D and RNA was isolated at 0 h, 1 h and 2 h. (H) RNA immunoprecipitation (RIP) assays were performed using NF90 or NF45 antibodies. Specific primers were used to detect HIF-1α or GAPDH, and RIP enrichment was determined relative to an input control. (I) Knockdown of DANCR decreased the interaction of HIF-1α with NF90, as detected by RIP assay using NF90 antibody. Data are presented as mean ± SD; Student's t-tests; *P<0.05. The experiments were independently repeated 3 times.