2 phenylethynesulfonamide pes

For induction and inhibition of Hsp70 expression or function, certain chemicals were utilized, including the HSF1 inducer U-133, which was previously selected from the library of small molecules of the Pacific Institute of Bioorganic Chemistry, Russian Academy of Sciences (Vladivostok, Russia). U-133 is an acetylated tris-O-glucoside echinochrome and it was obtained by the chemical modification of the sea urchins pigment echinochrome [31 ]. U-133 was found to induce synthesis in human U-937 promonocytes [24 (link)] and in the substantia nigra of rats [25 (link)]. The HSF1 inhibitor CL-43 was previously shown to inhibit Hsp70 synthesis in various cancer cells [26 (link)]. Moreover, 2-phenylethynesulfonamide (PES) or pifithrin-μ, an inhibitor of the chaperonic function of Hsp70 [32 (link)], was purchased at Sigma-Aldrich (St. Louis, MO, USA).

Oblongifolin C (>95% purity) was isolated from the fruits of Garcinia yunnanensis Hu using the same preparation method as previously described (Wang et al., 2016 (link); Supplementary Figures S1–S4; Table S1). The plants were collected at Dehong, Yunnan, People’s Republic of China in August 2015 and identified by Dr Guo-Dong Li (Yunnan University of Traditional Medicine) by referring to the voucher specimen (KUN 0941149) that was deposited at the Kunming Institute of Botany, Chinese Academy of Science. We procured 2-Phenylethynesulfonamide (PES, >95% purity) from Sigma-Aldrich (Cat # P0122-5MG) and Cisplatin from Selleck (Cat # S1166, >99% purity). They were, respectively, dissolved in dimethyl sulfoxide (DMSO) and dimethylformamide (DMF) to produce a stock concentration of 10 mM and were stored at −20°C. We purchased a FITC Annexin V PE Apoptosis kit from BD Biosciences (Cat # 556,547), Cell Counting Kit-8 (CCK-8) from Dojindo (Cat # CK04) and 4′,6-diamidino-2-phenylindole hydrochloride (DAPI) from Sigma-Aldrich (Cat #D9542). We purchased recombinant protein Lysozyme C (Cat #: CI91), Alpha-enolase (Cat #: CR49), Caspase-14 (Cat #: CG19) from Novoprotein company (Shanghai, China). HSPA2 (Cat #: H00003306-P01) from Amyjet Scientific company (Shanghai, China), Histone deacetylase 4 (HDAC4, Cat #: 11807-HNCB) from Sino Biological (Beijing, China).