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. 2021 Jul 1;22(13):7123. doi: 10.3390/ijms22137123

Figure 5.

Figure 5

MDIVI-1 treatment decreases the level of apoptotic Bax at mitochondria. (A–H) Detection of Bax and mitochondria by CLSM examinations in CS3BE-wtCSA and CS3BE cells. (A,B) untreated; (C,D) MDIVI-1 (10 µM for 24 h); (E,F) CCCP (20 µM for 16 h); (G,H) MDVI-1 and CCCP treated. Living cells were stained with Mitotracker® Deep Red FM (detected in red), fixed, permeabilized and labelled with polyclonal anti-Bax Ab (green). Nuclei are reported in blue (DAPI). Colocalization areas (detected in yellow) are shown in merged images. Insets represent separate channel images. For CS3BE cells a higher-power magnification image of a selected cell for each experimental condition is shown, with arrows indicating Bax-positive mitochondria. Scale bars, 20 µm. Panels are representative of three independent experiments. (I). Mean fluorescence value of cytoplasmic Bax in each experimental condition. (L). Quantification of colocalization of Bax with mitochondria after treatment with MDIVI-1 and/or CCCP, as determined by Pearson’s correlation coefficient measured in all microscopy images (mean ± SD). *** p< 0.0001, ** p < 0.01 by Mann–Whitney test.