Extended Data Fig. 3: Human phospho-kinase antibody array analysis and phosphoproteomic analysis of HeLa cells upon HD5 treatment.
a, Human phospho-kinase antibody array analysis of the phosphorylation of major kinases in the absence and presence of 4 μM HD5 as instructed by the manufacturer. Significance compared with solvent control (0 μM) group was calculated using two-tailed unpaired Student’ s t-test, and indicated as the p value. ns: no signification, **p < 0.01, ***p < 0.001 and ****p < 0.0001. Data are presented as mean±s.d. b, Top 50 proteins in phosphoproteomic analysis phosphorylated upon HD5 stimulation and dephosphorylated upon suramin treatment. Many of those proteins are involved in cytoskeleton related pathway such as Ras signaling pathway (RALGAPA1), Focal adhesion (CAV2), tight junction (CTTN), PI3K-Akt signaling pathway (PKN2), intermediate-sized filaments formation (KRT8), MAPK signaling (MAPK1), determining cell morphology (MISP), Rap1 interacting factor 1(RIF1), microtubule motor activity (KIF4B),cell junction organization (PLEC) and GTPase activator activity (USP6NL). c, GO terms in cellular component (CC) ontology of the phosphoproteome altered by HD5. A total of 363 phosphorylated sites, including 316 pSer (pS), 30 pThr (pT), and 17pTyr (pY) were selected as high-confidence targets from phosphoproteomic analysis for subsequent gene ontology (GO) enrichment analysis. Results are representative of three independent experiments.
