Fig. 2: Enhancement of specific GBM cell inhibition by HM-NPs@G.

a Flow cytometry analysis of U87MG cell targeting following 6 h incubation with Cy5 loaded HM-NPs, CM-NPs, MM-NPs or NPs (Cy5 concentration was 10 μg/mL). The flow cytometry analyses were representative data from three independent experiments. CLSM images (b) and (c) co-localization analysis of U87MG cells incubated with HM-NPs, CM-NPs, MM-NPs or NPs for 6 h. (Mitochondria stained by Mito tracker Red (Red), Nuclei stained by Hoechst (blue), FITC-labeled NPs (green)). The concentration of FITC was 10 μg/mL. Scale bar = 10 µm. The CLSM images and co-localization analysis were representative data from three independent experiments. d U87MG cell viability after 72 h incubation with free Gboxin, NPs@G, CM-NPs@G, MM-NPs@G, or HM-NPs@G (n = 4 biologically independent samples). Data are presented as mean ± SD (one-way ANOVA and Tukey’s multiple comparison test). e IC₅₀ values in normal cells (N2a, BV2, HA1800 and hCMEC/D3 cells) and GBM cells (U87MG, U251, U251-TR and X01) after 72 h incubation with HM-NPs@G or free Gboxin (n = 4 or 5 biologically independent samples, exact n seen in Supplementary Fig. 9). f ATP concentrations in U87MG cells treated with free Gboxin, NPs@G, CM-NPs@G, MM-NPs@G, or HM-NPs@G for 72 h (n = 3 biologically independent samples). Data are presented as mean ± SD (one-way ANOVA and Tukey’s multiple comparison test). g Quantitative analysis of JC-1 monomer fluorescence intensity in U87MG cells treated with free Gboxin, NPs@G, CM-NPs@G, MM-NPs@G, or HM-NPs@G) for 72 h (n = 4 biologically independent samples). Data are presented as mean ± SD (one-way ANOVA and Tukey’s multiple comparison test). h Schematic illustration of JC-1 structure changes resulting from changes in mitochondrial membrane potential changes. i TEM images of mitochondria in U87MG cells treated with PBS, free Gboxin, CM-NPs@G, MM-NPs@G or HM-NPs@G. The red arrows indicated the damaged mitochondria structure after treatment of nanomedicines. The TEM images were representative data from three independent experiments. j Western blotting analysis of cytochrome c (Cyto C) and apoptosis-related proteins treated with free Gboxin, NPs@G, CM-NPs@G, MM-NPs@G, or HM-NPs@G) for 72 h. The immunoblots were representative data from three independent experiments. k Schematic illustration of the mechanism of HM-NPs@G mediated GBM cell apoptosis. For Fig. 2d and Fig. 2f–j, the concentration of Gboxin was 800 nM. Source data are provided as a Source Data file.