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. 2023 Jan 22;10(8):2205668. doi: 10.1002/advs.202205668

Figure 3.

Figure 3

CircWDR37 stimulates NF‐κB‐mediated chemotherapy‐induced proinflammatory SASP genes transcription. a) GSEA analysis of the RNA‐seq results of si‐circWDR37 and siSCR transfected S18 cells. NES, normalized enrichment score; FDR, false discovery rate. b) RT‐qPCR results for the mRNA expression of the indicated NF‐κB‐targeted genes in HONE1 cells with circWDR37 knockdown. c) Luciferase assay of NF‐κB activity in HONE1, S18 and HK1 cells transfected with si‐circWDR37 or siSCR. d) RT‐qPCR results for the mRNA expression of the SASP component in si‐circWDR37 or siSCR transfected HONE1 cells with cisplatin or gemcitabine treated for 24 h. e) Western blots analysis of cyclin D1 in si‐circWDR37 or siSCR transfected HONE1, S18, and HK1 cells with cisplatin or gemcitabine treated for 24 h. f) Western blots analysis of indicated proteins in HONE1, S18, and HK1 cells treated with si‐circWDR37or siSCR. g) Western blots analysis of the indicated proteins in cytoplasmic (cyto) and nuclear (neu) fractions of HONE1 and S18 cells transfected with si‐circWDR37 or siSCR. Lamin B1 and α‐tubulin were used as nuclear and cytoplasmic markers, respectively. h) Fluorescence microscopy analysis of HONE1 and S18 cells transfected with si‐circWDR37 or siSCR, p65 nuclear fluorescence intensity was determined by ImageJ software. Nuclei were stained with DAPI (blue). Scale bar, 20 µm. Mean (n = 3) ± s.d. (Data were analyzed by (b–d) one‐way ANOVA with Dunnett's post‐hoc test and (h) one‐way ANOVA with Scheffe's post‐hoc test). p‐value < 0.05 indicates statistical significance. N.S. indicates no significance.