Fig. 2

Cellular uptake of CeONZs by hESCs and hESC-CMs. A Immunofluorescence analysis revealed the expression of pluripotency markers (OCT4, SSEA-4) in H9 hESCs and CM markers (cTNT, α-Actinin) in H9-CMs (Scale bar = 50 μm). B Flow cytometric analysis of the percent of cTNT⁺ CMs on differentiation day 30. The gray curve represents the blank control without staining, the green curve represents the negative control sample stained with the secondary antibody only, and the blue and red curves represent the H9-CMs stained with anti-cTNT antibody before and after purification respectively. C, D Cellular uptake quantified by cerium concentration per 10,000 cells in H9-CMs (C) and H9 hESCs (D) exposed to 60 μM and 6 μM CeONZs, respectively, at different time points. E CeONZs persistency in H9-CMs, quantified by cerium concentration per 10,000 cells after 24 h exposure to 60 μM CeONZs and further incubation with plain RB medium at days 3 (D3), and 5 (D5) after CeONZs removal. F CeONZs persistency in H9 hESCs after 24 h exposure to 6 μM CeONZs and further incubation with plain E8 at D3 and D5 after CeONZs removal. G Cerium concentration in mitochondria per 10,000 cells exposed to CeONZs (60 μM) in H9 hESCs and H9-CMs. Ctrl represents H9 hESCs, and H9-CMs without CeONZs treatment. H Statistically counted relative cerium concentration in H9 hESCs and H9-CMs co-incubated without drug treatment (Ctrl), with 2-DG and NaN₃, Genistein, and 2-DG, NaN₃, together with Genistein, before being exposed to 60 μM CeONZs for 2 h. Data are represented by means ± SEM (n ≥ 3) and statistical significance was determined by one-way ANOVA with a Tukey post-test. ns means no significance, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001