Pcs 100 042

Human vascular smooth muscle cells (hVSMC; primary aortic smooth muscle cells, ATCC-PCS-100-012) and HEK 293 T cells (ATCC-CRL-11268) were purchased from American Type Culture Collection. hVSMC were cultured in vascular basal media (ATCC PCS-100-030) supplemented with the vascular smooth muscle growth kit (ATCC PCS-100-042), 100 U/mL penicillin, 100 μg/mL streptomycin. hVSMCs were used between passages 2–5. HEK 293 T cells were cultured in high-glucose Dulbecco’s Modified’s Eagle Medium containing 10 mg/mL sodium pyruvate, 2 mM L-glutamine, 10% FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin. The HTLA cell line, a HEK293 cell line stably expressing a tTA-dependent luciferase reporter and a β-arrestin2-TEV fusion gene^{18 (link)}, was generously provided by the laboratory of Dr. Bryan Roth and maintained in high glucose Dulbecco’s Modified’s Eagle Medium supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, 100 µg/mL hygromycin B, and 2 µg/mL puromycin. All cells were cultured in a humidified environment at 37 °C, 5% CO₂.

The primary HASMCs were purchased from the American Type Culture Collection (ATCC PCS-100-012, Rockville, MD, USA). HASMCs were cultured in the vascular cell basal medium (ATCC PCS-100-030) with a vascular smooth muscle cell growth kit (ATCC PCS-100-042) at 37 °C in a 5% CO₂ incubator. The HASMC passage number in the experiments was P7–P9. In the treatment group, HASMCs were pretreated with NAC prior to PCS administration. NAC was always present during PCS stimulation.

hVSMCs (primary aortic smooth muscle cells, ATCC PCS-100-012) were purchased from American Type Culture Collection. As described previously [24 (link), 25 (link)], hVSMCs were cultured in vascular basal media (ATCC PCS-100-030) supplemented with the vascular smooth muscle growth kit (ATCC PCS-100-042), containing 100 U/mL penicillin, and 100 μg/mL streptomycin. hVSMCs were used between passages 2–5.