Journal: Theranostics

Article Title: Vine-inspired zinc-ion modified black phosphorus coating accelerates bone tissue infiltration of 3D printed scaffolds

doi: 10.7150/thno.113623

Figure Lengend Snippet: Zn@BP/Si activated YME1L to reduce abnormal mitochondrial fission to regulate mitochondrial dynamics. (A-B) GO and KEGG enrichment analyses revealed enrichment for biological processes. (C) Representative fluorescence images of FITC-labeled Zn@BP/Si endocytosis after Zn@BP/Si treatment for 10 min. Lysosomes were labeled with Lamp1. (D) Destination of Zn 2+ in mitochondria after Zn@BP/Si exposures. (E) Mito SOX detected mtROS content and (F) JC-1 detected mitochondrial membrane potential of HUVECs to reveal mitochondrial function. (G) Western blotting detected expressions of YME1L, proteins related to mitochondrial fusion and division, and apoptosis. (H) Mito-tracker staining showed the morphological changes of mitochondria after zinc chelator TPEN was added. (I) Cells stained with the Annexin-V/PI dye and the apoptosis rates were analyzed by flow cytometry. * P < 0.05; ** P < 0.01; *** P < 0.001. (J) The schematic diagram of regulation mitochondrial dynamics by upregulating YME1L activity via Zn@BP/Si.

Article Snippet: After blocking with BSA, the membranes were incubated overnight at 4 ºC with primary antibodies [anti-Bax (Abcam, 1:5000); anti-Bcl-2 (Abcam, 1:2000); anti-caspase-3 (Abcam, 1:2000); anti-cleaved caspase-3 (Abcam, 1:5000); anti-YME1L (Proteintech, 1:1000); anti-Drp1 (Abcam, 1:1000); anti-Mfn2 (Proteintech, 1:20000); anti-OPA1 (Abcam, 1:2000); anti-PRELID1 (Abcam, 1:500); anti-TIMM17A (Proteintech, 1:20000); anti-β-actin (Proteintech, 1:500000)].

Techniques: Fluorescence, Labeling, Membrane, Western Blot, Staining, Flow Cytometry, Activity Assay