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. 2023 May 1;133(9):e161929. doi: 10.1172/JCI161929

Figure 4. PD-L1 upregulation caused by Aurora A inhibition depends on STING/NF-κB activation.

Figure 4

(A and B) RNA-Seq analysis was performed on BxPC3 cells after treatment for 72 hours with DMSO or 1 μmol/L alisertib. (A) KEGG pathway analysis of genes differentially expressed between the DMSO- and alisertib-treated groups. The most substantially enriched pathways are shown. p.adjust, adjusted P value. (B) Heatmap of gene expression levels of the indicated cytokines or chemokines in DMSO- or alisertib-treated BxPC3 cells. (C and D) BxPC3 cells were pretreated for 6 hours with 10 μmol/L TPCA-1 or with 5 μmol/L BAY11-7082, followed by treatment for 72 hours with 1 μmol/L alisertib, and PD-L1 expression was assessed by Western blotting (C) and qRT-PCR (D). (E and F) qRT-PCR analysis of IFNB expression in BxPC3 cells after the indicated concentrations (E) and durations (F) of alisertib treatment (n = 3). (G) qRT-PCR analysis of IFNB expression in BxPC3 cells after treatment with the indicated siRNA (n = 3). (H) qRT-PCR analysis of IFNB expression (n = 3). (I–K) WT BxPC3 cells or STING–/– BxPC3 cells were treated with 1 μmol/L alisertib for 72 hours. (I) Western blot analysis of PD-L1 and STING protein levels. qRT-PCR analysis of PDL1 (J) and IFNB (K) mRNA levels (n = 3). Data indicate the mean ± SD. ***P < 0.001 and ****P < 0.0001, by 1-way ANOVA (D–H) and 2-way ANOVA (J and K).