Fig. 4
SOR@TF-Fe3+ NVs induced ferroptosis in HCC cells. A qPCR was used to analyze the expression of ACSL4 mRNA. B Western blot analysis of COX2 protein expression. C The laser confocal image showed the ROS production after treatment for 24 h. Scale bar: 10 μm. D The content of MDA after treatment for 24 h was detected by the kit. E Flow cytometry analysis was used to test the change of ΔΨm. On the right is the histogram of curve migration quantitative analysis. F CCK-8 was used to detect the activity changes of HCC cells after 24 h in each treatment group. G The cell clone formation of SMMC7721, HepG2 and HepG2-SR cells in each experimental group was detected after 7 days of treatment, Dosage: SOR 1 μg/mL, nanovesicles 10 μg/mL (protein weight). H Cell growth was measured by clone formation assay when co-incubated with ferroptosis inhibitor Ferrostatin-1 (Fer-1, 2 μM), necrosis inhibitor Necrostatin 1 (Nec, 0.5 μM) and apoptosis inhibitor Z-VAD-FMK (Z-VAD, 2 μM). And the quantitative analysis of clone formation was shown on the right. Data are expressed as mean ± standard error (SEM), n = 3, ns: no significant, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, one-way analysis of variance, ANOVA