Fig. 3: circMAP3K13 harbors an active IRES sequence, and can be translated into a novel protein. | Cell Death & Disease

Fig. 3: circMAP3K13 harbors an active IRES sequence, and can be translated into a novel protein.

From: MAP3K13-232aa encoded by circMAP3K13 enhances cisplatin-induced pyroptosis by directly binding to IKKα in gastric adenocarcinoma

Fig. 3

A Schematic of circMAP3K13, which consists of 803 nucleotides and is predicted to encode a novel protein of approximately 26 kDa. B Schematic representation of plasmid constructs. The circMAP3K13-FLAG plasmid overexpresses circMAP3K13 with a 3×FLAG tag. The downstream flanking sequence deletion mutant lacks the downstream complementary sequence required for circularization. The ATG-mutation construct disrupts the start codon, altering the open reading frame. The linear-MAP3K13-232aa-FLAG plasmid expresses the same ORF of circMAP3K13 in a linear form. C Western blot analysis of circMAP3K13-FLAG and linear-MAP3K13-232aa-FLAG. D RNAfold software prediction of the secondary structure of circMAP3K13 and a putative internal ribosome entry site (IRES). E Schematic of five recombinant plasmids generated to assess IRES activity by dual-luciferase reporter assay. F Relative luciferase activity of plasmids 1–6. G AlphaFold3–based 3D structure prediction of MAP3K13-232aa. The MAP3K13-232aa sequence is highlighted in red within the full MAP3K13 protein (left) and the specific MAP3K13-232aa peptide is shown in cyan (right). Unless otherwise specified, statistical analysis was performed using a two-tailed Student’s t-test. **P < 0.01; ***P < 0.001.

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