Additional file 3 of M2 macrophage-secreted KYNU promotes stemness remodeling and malignant behavior in endometrial cancer via the SOD2-mtROS-ERO1α-UPRER axis

Supplementary Material 3: Fig S3. Cell proliferative capacity A, cell invasion B, and cell apoptosis rate C of HEC-1A cells cocultured with M1 or M2 macrophages were detected by a CCK-8 assay, a transwell assay and flow cytometry. D IC50 values for the KYNU inhibitor in HEC-1A cells were detected by a CCK-8 assay. The concentrations used were 50 μmol/L, 100 μmol/L, 500 μmol/L, and 1,000 μmol/L. E The interaction between KYNU and SOD2 was verified by Co-IP. F The regulatory effects of the KYNU inhibitor, recombinant KYNU and 3-HAA on SOD2 were confirmed by western blot. G Flow cytometry analysis of the influence of KYNU, SOD2, MitoTempol and their combination on mtROS levels. The proliferative capacity H, invasion I, and apoptosis rate J of HEC-1A cells pretreated with the KYNU inhibitor or mitoTempol were detected by a CCK-8 assay, a transwell assay and flow cytometry. K western blot analysis of the influence of the KYNU inhibitors 3-HAA and mitoTempol as well as their combination on ERO1α expression. The cell proliferative capacity L, invasion M, and apoptosis rate N of the HEC-1A cells pretreated with the KYNU inhibitor or ERO1α-OE were detected via the CCK-8 assay, transwell assay and flow cytometry. O Two predicted binding sites of the KYNU promoter region with the transcription factor ATF4 were verified by a ChIP assay in the HEC-1A cell line. The data are presented as the means ± SDs (n = 3 per group), Student’s t test, one-way analysis of variance (ANOVA). *P < 0.05, **P < 0.01, ***P < 0.001.