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

Figure 5. Aurora A inhibition–induced PD-L1 expression is mediated by cGAS dephosphorylation.

Figure 5

(A–C) WT BxPC3 cells or CGAS–/– BxPC3 cells were treated with 1 μmol/L alisertib for 72 hours. (A) Western blot analysis of PD-L1 and cGAS protein levels. PD-L1 and cGAS were detected separately in 2 gels using the same biological samples, and GAPDH in each gel served as the loading control. qRT-PCR analysis of PDL1 (B) and IFNB (C) mRNA levels (n = 3). (D) Co-immunoprecipitation of Aurora A and cGAS. HEK293T cells were transfected with the indicated vectors encoding HA–Aurora A and Flag-cGAS. Whole-cell lysates were immunoprecipitated with anti-Flag beads, and the interactions were analyzed by Western blotting. (E) BxPC3 cells were synchronized with 10 μmol/L Ro-3306 for 16 hours and released into mitosis in the presence of alisertib at the indicated concentrations. Phosphorylation of cGAS was analyzed by Phos-tag electrophoresis. Data indicate the mean ± SD. ****P < 0.0001, by 2-way ANOVA.