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. Author manuscript; available in PMC: 2020 Jun 4.
Published in final edited form as: J Am Coll Cardiol. 2019 Jun 4;73(21):2722–2737. doi: 10.1016/j.jacc.2019.03.476

Figure 5. Effect of conditioned media from iPSC-ECs treated with e-liquid on macrophage polarization.

Figure 5.

(A) Conditioned media was collected from iPSC-ECs after 48 hours of incubation with each e-liquid flavor, and then exposed to macrophage-like cells for 48 hours. Phenotype of macrophages was analyzed by flow cytometry for M1 (CD40) and M2 (CD163) markers. (B) After 48 hours incubation with conditioned media, the media was replaced with the fresh macrophage culture media for 16 hours to determine cytokine expression and ROS levels. Expression of cytokines produced by macrophages was determined by quantitative real-time RT-PCR. (C) Intracellular ROS levels produced by macrophages was determined using a ROS-Glo H2O2 assay. Data are represented as mean ± SEM. Statistically significant from controls (Bonferroni-adjusted P<0.05). MAR0: Marcado e-liquid without nicotine; MAR18: Marcado e-liquid with 18 mg/ml of nicotine; RY0: RY4 e-liquid without nicotine; RY18: RY4 e-liquid with 18 mg/ml of nicotine.