Fig. 3: LINC00239 interacts with Keap1. | Cell Death & Disease

Fig. 3: LINC00239 interacts with Keap1.

From: Long noncoding RNA LINC00239 inhibits ferroptosis in colorectal cancer by binding to Keap1 to stabilize Nrf2

Fig. 3

A Proteomic approach to identify the LINC00239-specific interactors. B Proteome-wide accurate quantification and significance. C Western blot of the proteins from anti-sense LINC00239 and LINC00239 pull-down assays. D RNA immunoprecipitation experiments were performed using anti-Keap1 antibody, and specific primers were used to detect LINC00239. E ImmunoFISH images of LINC00239 and Keap1 in SW620 cells. F Top, the predicted secondary structure of LINC00239. Bottom, the in vitro-transcribed full-length LINC00239 and deletion fragments with the correct sizes indicated. G Deletion mapping of the Keap1-binding domain in LINC00239. Top, diagrams of full-length LINC00239 and the deletion fragments. Bottom, immunoblot analysis for Keap1 in the protein samples pulled down by different LINC00239 constructs. H The RNA Pull-down analysis of FLAG-tagged Keap1 versus domain truncation mutants) retrieved by in vitro-transcribed biotinylated LINC00239. I RIP assays show the association of the Kelch domain with LINC00239. Data shown represent mean ± SD from three independent experiments. ns P > 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, Student’s t test.

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