Phospho-TACC3 (Ser558) Antibody - #AF4506

Figure 3 Characterization of cellular response to PROTACs in AML cells. A) KG1A and Kasumi‐1 cells were stained with a carboxyfluorescein succinimidyl amino ester (CFSE) probe and cultured with AURKA PROTACs (1 × 10−6 m) for 72 h. CFSE fluorescence was analyzed by flow cytometry. MFI, mean fluorescence intensity. B) KG1A cells were treated with AURKA PROTACs (1 × 10−6 m) for 48 h. The cell cycle profile was assayed by FACS with propidium iodide (PI) staining. The cell cycle phase distribution was analyzed by FlowJo software. C) A heatmap of the relative normalized abundance of proteins in TMT‐based quantitative proteomic assays. D) TMT‐based quantitative proteomics after treatment with PROTACs (500 × 10−9 m) or the DMSO Vehicle for 6 h in KG1A cells. The top 100 decreased proteins were subjected to g:Profiler to perform gene ontology (GO) analysis. E) Gene set enrichment analysis (GSEA) of RNA‐seq data from KG1A cells treated with AURKA PROTACs (1 × 10−6 m) for 48 h. F) KG1A cells were treated with AURKA PROTACs (1 × 10−6 m) or ATRA (0.6 × 10−6 m) for 48 h. Cell surface CD34 expression was analyzed by FACS. MFI, mean fluorescence intensity. G) KG1A cells were arrested with nocodazole (0.1 µg mL−1) for 16 h followed by PROTACs or MLN8237 treatment with indicated concentration for 6 h. AURKA localization were detected by immunofluorescence. Scale bar: 5 µm. H) KG1A cells were arrested with nocodazole (0.1 µg mL−1) for 16 h followed by DMSO, PROTACs (1 × 10−6 m) or MLN8237 (50 × 10−9 m) treatment for indicated times. The expression of AURKA and related proteins were detected. Statistics, significance: one‐way ANOVA with Bonferroni correction (A,F); ns, not significant;