Fig 2: AGAP2‐AS1 was functionally related to EIF4A3 in activating MRC‐5 cells. (a) CCK‐8 assay was used to assess the viability of MRC‐5 cells. (b) and (c) Migrated and invaded MRC‐5 cells analyzed by transwell assay were counted after the indicated treatments. (d) and (e) Wound healing rates were calculated after 24 h of culture. MRC‐5 cells were transfected with the referred vectors. (f) mRNA expression levels of the listed cytokines in MRC‐5 cells underwent the indicated transfections. (g) The protein expression levels of fibroblast activating protein and α‐smooth muscle actin in MRC‐5 cells transfected with the indicated vectors. Three independent experiments were performed for each assay. *p < 0.05, **p < 0.01

Fig 3: Circ_0020256 recruited EIF4A3 protein to stabilize KLF4 mRNA.A Circular RNA Interactome database analysis of the potential binding between circ_0020256 and EIF4A3 protein. Circ_0020256 expression in CCA specimens (B) and cell lines (C) was detected by qRT-PCR. D Schematic diagram of circ_0020256 formation and Sanger sequencing was used for identifying circ_0020256. The expression of circ_0020256 and linear NSMCE4A mRNA after exposure to RNase R (E) or actinomycin D (F) were assessed by qRT-PCR. The interaction between circ_0020256 and EIF4A3 protein was verified by RIP (G) and RNA pull-down assay (H). I qRT-PCR analysis of circ_0020256 level in CCA cells after transfection with OE-circ_0020256 or sh-circ_0020256. J EIF4A3 expression in CCA cells with different treatments was detected by western blotting. K The positive correlation between circ_0020256 and EIF4A3 expression in CCA samples. L The stability of KLF4 mRNA after exposure to actinomycin D was evaluated by qRT-PCR. M The interaction between KLF4 mRNA and EIF4A3 protein in circ_0020256-silenced CCA cells was determined by RIP assay. 45 paired patient samples were detected in B and K. *P < 0.05, **P < 0.01, and ***P < 0.001.

Fig 4: Exosomal circCOL1A1 promotes angiogenesis through Smad2/3 pathway. HUVECs were transfected with pcDNA3.1-NC or pcDNA3.1-EIF4A3, followed by treatment with CRC cell-derived exosomes. (A) The protein levels of VEGFR and p-Smad2, Smad2, p-Smad3, and Smad3 were detected by western blot. *, P < 0.05, **, P < 0.01, ***, P < 0.001

Fig 5: Hypothesis of CDR2L and CDR2 involvement in protein synthesis in Purkinje neurons. CDR2 localizes to the nucleus and directly interacts with nuclear speckle protein eIF4A3. eIF4A3, in conjugation with other cytoplasmic initiation factors, facilitates mRNA binding to the 40S ribosomal subunit. This event is important for mRNA maturation and translation, ultimately resulting in the synthesis of new proteins. CDR2L interacts with ribosomal subunit protein rpS6; therefore, we propose that CDR2L and CDR2 are both involved in the process of protein synthesis. Furthermore, Yo antibody (green) binding to CDR2L in Purkinje neurons of PCD patients may, therefore, interfere with the function of the ribosomal machinery, resulting in disrupted mRNA translation and/or protein synthesis.