Fig. 5.

CircPHLPP2 facilitates IL36γ transcription by binding to ILF3. a The co-localization assay of circPHLPP2 with ILF3 or PABPC1 in HCT116 cells was identified by RNA-FISH and immunofluorescence, scale bar = 10 μm. b Binding of circPHLPP2 with ILF3 was identified by RNA pull-down analysis using the MS2-tagging system. c RIP assay with an anti-ILF3 antibody or control IgG was performed in HCT116 and SW480 cells, followed by RT-qPCR to assess the enrichment of circPHLPP2. d Mass spectrometry analysis identified ILF3. e Western blotting analysis of cytoplasmic and nuclear ILF3 protein in HCT116 and SW480 cells with or without circPHLPP2 overexpression. f Coimmunoprecipitation (Co-IP) and western blot analysis of the interaction between ILF3 and exportin-5 in circPHLPP2 overexpression and control HCT116 cells. g Relative expression of IL36γ was measured by RT-qPCR in HCT116 and SW480 cells transfected with si-ILF3#1, si-ILF3#2 or si-NC. h Relative luciferase activity (FL/RL) was measured in HCT116 and SW480 cells after co-transfection of the IL36γ luciferase construct with ILF3 overexpression or control vector. i-j Relative expression of IL36γ was measured by RT-qPCR in HCT116 (i) and SW480 (j) cells with circPHLPP2 overexpression or ILF3 knockdown. Data in c and h were calculated by Student’s t test. Data in g and i-j were calculated by one‑way ANOVA test. ***P < 0.001, ****P < 0.0001