Phospho-Nucleophosmin (Thr95) Antibody - #AF2372

Fig. 5: AURKA promoted ES cell apoptosis and ferroptosis resistance via phosphorylating Thr95 of NPM1. A Silver staining of co-immunoprecipitation with AURKA antibody in A673. B Results of protein mass spectrometry revealed the physical interaction of AURKA and NPM1. C Results of immunoprecipitation to confirm the interaction relationship of AURKA and NPM1 in ES cells. The corresponding original western blots are shown in Fig. S9. D WB analysis identified the expression level changes of p-NPM1 at Ser4, Thr95, Ser125, Thr199, and the total NPM1 in ES cell lines after treatment with TCS7010. The corresponding original western blots are shown in Fig. S10. E WB analysis identified the expression level changes of p-NPM1 at Ser4, Thr95, Ser125, Thr199, and the total NPM1 in ES cell lines after AURKA knockdown. The corresponding original western blots are shown in Fig. S10. F Colony formation assay showed the reproductive ability of ES cells with NPM1 knockdown. G Colony formation assay determined the reproductive ability of ES cells with NPM1 knockdown in the absence or presence of indicated cell death inhibitors (10 µM ZVAD-FMK and 10 µM ferrostatin-1). H Columnar statistical chart based on the CCK-8 assays indicated the cell viability after NPM1 knockdown in the absence or presence of indicated cell death inhibitors (10 µM ZVAD-FMK and 10 µM ferrostatin-1) for 48 h. I Columnar statistical chart indicated changes in the apoptosis rates of ES cell lines after NPM1 inhibition with siRNA. J Detection of the intracellular ROS levels in ES cells after NPM1 inhibition. K Detection of the intracellular lipid ROS levels in ES cells after NPM1 inhibition. L Changes of the relative intracellular Fe2+ levels in ES cells after NPM1 inhibition. M WB analysis indicated changes in the apoptosis-related gene markers (PARP, Bcl2, Bax) and the ferroptosis-related marker GPX4 after NPM1 inhibition in ES cells. The corresponding original western blots are shown in Fig. S11. Values represented the mean ± SD from 3 independent experiments.