Additional file 1: of Identification of a novel S6K1 inhibitor, rosmarinic acid methyl ester, for treating cisplatin-resistant cervical cancer

Figure S1. Chemical structures of high-ranking virtual screening hits from both docking- and similarity-based method. Figure S2. RAME inhibits H2B phosphorylation by S6K1 in vitro. In vitro kinase assay with RAME was performed in a dose dependent manner using recombinant H2B, active S6K1, and cold-ATP. Figure S3. Effects of RAME on lung cancer cell lines. Immunoblotting analysis of A549 and H1299 cells treated with RAME (40, 80 μM) for 24 h. Figure S4. Effects of RAME and PF-4708671 on phosphorylation of Akt. Immunoblotting analysis of HeLa cells treated with RAME (40 μM) or PF-4708671 (20 μM) for 24 h. Figure S5. RAME induces apoptosis in cervical cancer cells. (A) Immunoblotting analysis of HeLa cells treated with RAME (40 or 80 μM) for 24 h. (B) Flow cytometric analysis of HeLa cells treated with RAME (80 μM) for 24 h. Figure S6. RA does not enhance the effects of cisplatin in cervical cancer cells. (A) The mRNA levels of autophagy-related genes in SiHa cells treated with or without cisplatin (5 μM) and RA (80 μM) for 24 h. (B) The mRNA levels of apoptosis, DNA repair, and cell cycle arrest marker genes in SiHa cells treated with or without cisplatin (5 μM) and RA (80 μM) for 24 h. Error bars correspond to mean ± SEM (n = 3). *p