Fig. 3: HMPB-HKN15-induced ferroptosis in 4T1 cells in vitro. | Nature Communications

Fig. 3: HMPB-HKN15-induced ferroptosis in 4T1 cells in vitro.

From: A ferritin-targeted biohybrid triggering ferroptosis immunotherapy via activating endogenous iron and replenishing exogenous iron simultaneously

Fig. 3

a, b CLSM images and plot profile of HMPB-H and ferritin co-localization analysis in 4T1 cells. Scale bar: 20 µm. c The intracellular ROS levels were monitored using DCFH-DA. Scale bar: 50 µm. d Intracellular levels of GSH by CLSM. Scale bar: 50 µm. e GPX4 expression using western blotting. f LPO levels detected by Liperfluo probe. Scale bar: 50 µm. g, h LPO levels estimated by flow cytometric quantification. i, j Annexin V/PI staining by flow cytometric quantification. k Calcein/PI staining assay. Scale bar: 50 µm. l Intracellular oxygen levels by CLSM. Scale bar: 50 µM. m, n Cell viability and LPO levels in 4T1 cells treated with HMPB-H + L in the presence or absence of Fer-1. o, p Corresponding gray value analysis of GPX4 and SLC7A11 expression in 4T1 cells after different treatments. For (a, c–f, k, l), experiment was repeated three times independently with similar results. The data in (h, j, m–p) are presented as the mean ± SD (n = 3 independent experiments). Statistical differences were analyzed by one-way ANOVA with Tukey’s multiple comparisons test.

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