Extended Data Fig. 3: Chemotherapy enhances SDF1 expression via upregulating c-JUN.
a, SCENIC package was used to analyze ECs from breast cancer patients and ovarian cancer patients with or without chemotherapy. The activity score of each transcription factor (TF) regulon was calculated and ranked. Top 10 TF regulons are marked. b, The gene sets representing upregulated TF activity after chemotherapy in breast and ovarian cancer patients were identified and then overlapped with necroptosis pathway (GSE121149) to perform Venn diagram analysis. Top 10 genes were used in each gene set. c, The scatter plot showed the activity of c-JUN transcription factor in the ECs of breast and ovarian cancer patients from scRNA-seq data. The black horizontal line in the plot represents the average value of the two groups. d, Cut&Tag assay of SDF1 promoter was performed with anti-c-JUN antibody. Promoter binding ability was determined with qPCR (n = 5 biological replicates). e, c-JUN was overexpressed in HUVECs. SDF1 promoter activity was determined by luciferase reporter assay (n = 4 biological replicates). f, HUVECs stimulated by CBP were treated with increasing doses of T5224. SDF1 promoter activity was determined by luciferase reporter assay (n = 4 biological replicates). g-j, PARP1 or MLKL was silenced with shRNA in HUVECs and treated with CBP. The expression levels of SDF1 and c-JUN were measured with western blot (g, i) and quantified by ImageJ (h, j) (n = 6 biological replicates). k, Schema illustrating the mechanism by which c-JUN binds to the SDF1 promoter, leading to increased SDF1 expression in ECs post-chemotherapy. ****P < 0.0001 was calculated using two-sided unpaired Wilcoxon rank sum test for c or two-way ANOVA followed by Tukey’s test as post-hoc analysis for d, e, f, h and j.
