Fig. 4: EIF4E2 destabilizes CXCL10 mRNA via AU rich element of 3'UTR. | Nature Communications

Fig. 4: EIF4E2 destabilizes CXCL10 mRNA via AU rich element of 3'UTR.

From: PARP inhibitor radiosensitization enhances anti-PD-L1 immunotherapy through stabilizing chemokine mRNA in small cell lung cancer

Fig. 4

a Top: Schematic of the psicheck2-Rluc-CXCL10 3′UTR reporter. Left: Western blot against EIF4E2 in Ctrl or EIF4E2 KO HEK293 cells. Quantification of relative Renilla luciferase mRNA level (middle) and protein level (right) in EGFP KO control (Ctrl) or EIF4E2 KO HEK293 cells transfected with psicheck2 or psicheck2-CXCL10 3′UTR plasmids (n = 3 biological replicates for mRNA detection and n = 4 biological replicates for protein detection). b Top: Schematic of the GFP-CXCL10 3′UTR reporter. Left: Western blot against EIF4E2 in LacZ KO control (Ctrl) or EIF4E2 KO for SBC5. Right: Quantification of GFP intensity in Ctrl or EIF4E2 KO for SBC5 expressing GFP-CXCL10 3′UTR (n = 4 biological replicates). c Top: Schematic of the psicheck2-Rluc-CXCL10 3′UTR WT and 3′UTR △ARE reporter. Bottom: Quantification of relative Renilla luciferase mRNA stability in Ctrl or EIF4E2 KO HEK293 cells transfected with psicheck2-Rluc-CXCL10 3′UTR WT or △ARE reporter plasmid for 24 h and then treated with Dactinomycin (DACT) for 4 h (n = 3 biological replicates). d Top: Schematic of the CXCL10-ORF + 3′UTR WT or 3′UTR △ARE plasmid and the mRNA produced (Created in BioRender. Ran, X. (2025) https://BioRender.com/v38l510). (Left) Relative level of CXCL10 genomic DNA (n = 4 biological replicates), (Middle) CXCL10 mRNA stability (n = 3 biological replicates), and (Right) western blot and quantification of CXCL10 protein levels (from one of two independent experiments) in SBC5 expressing CXCL10-ORF + 3′UTR WT or 3′UTR △ARE e Left: Schematic of SBC5-3′UTR WT or SBC5-3′UTR △ARE tumor engraftment in PBMC humanized NSG mice (Created in BioRender. Ran, X. (2025) https://BioRender.com/e93s595). Middle: Representative flow cytometry data for total T cells. Right: Cumulative data for total T cell (CD3+CD45+) in tumors (n = 6) on day 35. For (a–d), data are presented as mean ± SD. For (a) data are analyzed by two-way ANOVA. For (b–d), data are analyzed by two-tailed, unpaired t test with an exception for (e) by two-tailed, paired t test. (p: **** < 0.0001 < *** < 0.001 < ** < 0.01 < * < 0.05). All exact p values are listed in the source data. Source data are provided as a Source Data file.

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