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. 2020 Sep 21;12(9):2709. doi: 10.3390/cancers12092709

Figure 2.

Figure 2

BTZ is not toxic to normal cells when bound to targeted MSNs. (A) RPMI-8226 (RPMI) and BJhTERT were treated or not (control) with MSN-FOL, FOL-MSN-BTZ and free BTZ for 1 h and processed for TUNEL assay after 36 h. Nuclei were counterstained with DAPI. Cells were photographed at 10× magnification, and apoptotic cells from triplicate experiments were counted using Image J software (graphs on the right). (*) p < 0.05 vs. control. (B) A duplicate set of cells was processed for TEM analysis (see Materials and Methods). Red arrows: apoptotic cells; green arrows: parthanatic cell death; yellow arrows: MSNs. Scale bars 5 μm, × 2000 magnification. (C–F) Molecular profile of BTZ-induced death in MM cells and in BJhTERT cells line. A third set of both cell lines was treated as in (A,B), and cytosolic and mitochondrial fractions (C,D) or total proteins (E,F) were extracted and subjected to WB analysis to assess the expression of the indicated apoptotic markers. β-Actin and GAPDH were used as loading control; COX IV: mitochondrial marker to assess fractionation quality. NS: Non-Specific bands.