Growth media

Human adult aortic VSMCs (passage 3–10) were purchased from Cell Applications Inc. (354-05a). VSMCs were grown in growth media (Cell Applications Inc.), prior to being washed with Earle’s Balanced Salt Solution (Thermo) and seeded in basal media (Cell Applications Inc.) onto 12 kPa hydrogels, 18 h prior to drug treatment. Standard VSMCs culture was performed as described previously (Ragnauth et al., 2010 (link); Warren et al., 2015 (link)).
Quiescent VSMCs were stimulated with either angiotensin II (0.01–100 µM) (Merck) or carbachol (0.01–100 µM) (Merck) for 30 min. For all other drug treatments, quiescent VSMCs were pretreated with the stated dose for 30 min, prior to co-treatment with angiotensin II (10 µM) for an additional 30 min. Please see Supplementary Table S1 for a list of compounds used in this study.

Human RMS cell lines, Rh3, Rh5, Rh18, Rh30, and Rh41, were obtained from Childhood Cancer Repository (http://www.cccells.org) and were cultured in RPMI 1640 (Thermo Fisher Scientific (Thermo), Waltham, MA, Cat. #11875-093) containing 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Cat. #26140-079) and 1% penicillin/streptomycin (Thermo, Cat. #15140-122). RD was obtained from ATCC, Manassas, VA (Cat. #CCL-136) and grown in Dulbecco's modified Eagle's media (DMEM) (Thermo, Cat. #11995-065) supplemented with 10% FBS and 1% penicillin/streptomycin. Primary HSMM cell lines were obtained from Lonza Inc., Morristown, NJ (Cat. #CC-2580) and cultured in growth media from Cell Applications, San Diego, CA (Cat. #151-500). SJ-GBM cell line was obtained from Dr. Susan Baker (St. Jude's Hospital) and was grown in a base medium of Iscove's modified Dulbecco's medium (IMDM, Thermo, Cat. #12440061) with 10% FBS, 1% penicillin/streptomycin, 4 mM L-glutamine (Thermo, Cat. #25030081) and 1x insulin, transferrin, and selenous acid (ITS, Corning Life Sciences, Bedford, MA, Cat. #41400045). PDX explant primary cell culture CF-1 was grown in RPMI 1640 containing 10% FBS and 1% penicillin/streptomycin. All cell cultures were maintained at 37°C and 5% CO2. STR analyses of the cell lines are shown in Supplementary Table 1.

Rat primary dermal fibroblasts (RDFs), growth media, and passaging solutions were purchased from Cell Applications (San Diego, CA, USA). Cells were maintained in Culture Complete Growth Medium and penicillin/streptomycin solution in a humidified incubator with 5% CO₂ at 37°C. To eliminate any possible side effects of growth factors, cells were cultured in serum-free medium for 24 h before treatment with ANT extract.