Fig. 2: NCA changes mitochondrial ultrastructure, disturbs mitochondrial membrane potential and increases ROS generation. | Cell Death Discovery

Fig. 2: NCA changes mitochondrial ultrastructure, disturbs mitochondrial membrane potential and increases ROS generation.

From: Neocarzilin A induces apoptosis and mitochondrial disturbance by targeting reticulon 4-mediated endoplasmic reticulum stress

Fig. 2

A Representative transmission electron micrographs of HeLa cells treated with NCA or DMSO as indicated (scale bar top row 5000 nm, bottom row 1000 nm, mi=mitochondria). B Quantification of 50 mitochondria per repetition of mitochondrial size, mean gray value, cristae density and shape factors circularity and aspect ratio. C, D Analysis of mitochondrial membrane potential via JC-1 staining and flow cytometry upon C, D NCA or DMSO treatment and D concomitant BAPTA application. CCCP served as positive control. E Mitochondrial calcium levels upon treatment with NCA or DMSO as indicated by using Rhod2-AM dye and flow cytometry. F, G Mitochondrial superoxide generation after NCA or DMSO treatment as indicated measured with the MitoSOXTM dye by F flow cytometry and G live cell imaging. F Antimycin A served as positive control. Representative images shown on the bottom panel, nuclei in blue, MitoSOX in green (scale bar 25 µm). Brightness was adjusted to improve visibility. Absolute values B–D or normalized data E–G are presented in B box and whiskers (min to max) or C–G bar graphs as mean ± SD, n = 3. Statistical significance was analyzed by B two-tailed unpaired Student’s t-test or C–G one-way ANOVA with Dunnett’s posttest compared to mean of DMSO control (ns not significant, *P < 0.05, **P < 0.01, ***P < 0.001).

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