U87mg

HDF (human dermal fibroblast) cells, MDA-MB-231 (human breast cancer cells) and U-87 MG (human brain glioblastoma-astrocytoma cell lines) were obtained from ATCC, Manassas, VA, USA. HDF and MDA-MB-231 cells were grown in Dulbecco’s Modified Eagle’s Medium (DMEM) medium whereas U-87 MG was grown in DMEM/F12 medium with 10% fetal bovine serum (HyClone™ Fetal Bovine Serum (U.S.), Characterized) and 100 units/mL of penicillin/streptomycin at 37 °C in a humidified atmosphere in 5% CO₂ incubator.
The HDF, MDA-MB-231 and U-87 MG cell lines were seeded into 96-well microtiter plates (Corning^®, Saint Louis, MO, USA) at a density of 1.2*10⁴, 1.0*10⁴ and 1.5*10⁴ cells/wells, respectively. After overnight incubation, cells were exposed to increased concentrations of the ethanolic extract of D. sericea Vahl (125, 625 and 1.25 mg/mL) in cell culture medium. After incubation times of 24, 48 and 72 h, medium was replaced by a new fresh one containing 0.5mg/mL of MTT. After 2 h, at 37°C in 5% CO₂ cells were dissolved in DMSO. Absorbance was read at 570 nm by a spectrophotometer plate reader. The data were expressed as absorbance relative to untreated cells in the same experiment and standardized to 100%. All data points were performed in triplicate and in, at least, three independent experiments.

Glioma cell lines, including A172, U87MG and U373MG, and the astrocyte cell line SVG-P12 were purchased from ATCC (Manassas, VA, USA) and cultured in Dulbecco’s Modified Eagle’s Medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (GE Healthcare Life Sciences, South Logan, UT, USA), 100 μg/ml penicillin and 100 μg/ml streptomycin (Thermo Fisher Scientific) at 37 °C in a 5% CO₂ incubator. For cell counting, the cells were pretreated with DHEA (Sigma-Aldrich, St Louis, MO, USA) for 48 h and then treated with TMZ (Sigma-Aldrich) for an additional 48 h. Abiraterone was purchased from Selleckchem (Houston, TX, USA).

The U87-MG (human glioblastoma, HTB-14) cells, PC3 (prostate carcinoma, CRL1435) cells, PPC1 (primary prostate carcinoma-1) cells were purchased from ATCC. Murine WT-GBM glioblastoma cells were a kind gift from Gabriele Bergers (UCSF, USA) and P3, P8, P13 stem cell-like, P22, NCH421K cells were a kind gift from Rolf Bjerkvig, (University of Bergen, Norway), and M21 melanoma cells were the gift of David Cheresh (USA). Cells and tumors were prepared as described previously^{19 –21 (link)}.
Athymic nude mice (Hsd/Athymic Fox1 nu Harlan) were purchased from Envigo (Netherlands) and maintained under standard housing conditions of the Animal Facility of the Institute of Biomedicine and Translational Medicine, University of Tartu (Tartu, Estonia). For orthotropic GBM tumor models, we used NCH421K, P13, and P3 stem cell-like, WT-GBM cells. The individual GBM cells around 2–3 × 10⁵ in 3 μL PBS were intracranially implanted into mice brain 2 mm right and 1 mm anterior to the bregma. U87-MG and PC3 subcutaneous models were induced by injecting 2–9 × 10⁶ cells in 100 µl PBS subcutaneously into the right flank of 11–15 week old male and female nude mice. All animal experiments were performed in accordance with the procedures approved by the Committee of Animal Experimentation of Estonian Ministry of Agriculture, permits #42 and #48.

Carboxymethylcellulose sodium salt (CMC, DS = 0.7, M_w = 250 kDa; viscosity = 735 cps at 2% in H₂O, at 25 ºC), L-cysteine hydrochloride (Cys, HSCH₂CH (NH₂)COOH · HCl, ≥ 98%), and doxorubicin hydrochloride (DOX, C₂₇H₂₉NO₁₁ ⋅ HCl, ≥ 98.0%) were purchased from Sigma-Aldrich (USA). KLA (LAKLAKKLAKLAK) and KLAR7 (RRRRRRRKLAKLAKKLAKLAK) peptides were purchased from GenScript (USA). All of the other materials used in this research were detailed in the Supplementary Material.
Human embryonic kidney cells (HEK 293T, American Type Culture Collection - ATCC CRL-1573) were provided by the Federal University of Minas Gerais/UFMG. Human brain likely glioblastoma cells (U-87 MG, ATCC HTB-14) were supplied from Brazilian Cell Repository (Banco de Células do Rio de Janeiro: BCRJ, Brazil; cell line authentication molecular technique, Short Tandem Repeat (STR) DNA; quality assurance validation by international standard NBR ISO/IEC 17025:2005).