Fig. 4: NCA treatment leads to ER-derived cytosolic vacuolization and calcium increase. | Cell Death Discovery

Fig. 4: NCA treatment leads to ER-derived cytosolic vacuolization and calcium increase.

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

Fig. 4

A Representative phase contrast images of HeLa cells treated with NCA or DMSO as indicated (scale bar 25 µm). White arrows indicate vacuoles. B Representative transmission electron micrographs of HeLa cells treated with NCA as indicated (scale bar left image 5000 nm, right image 1000 nm). Red arrow indicates nuclear membrane budding. C Analysis of cytosolic calcium by Cal520 AM flow cytometry assay. HeLa cells were loaded with the dye and treated with 10 µM NCA for the indicated times. Cal520 AM mean fluorescence was recorded. D Representative confocal images of HeLa cells transfected with DsRed2 plasmid, treated with 10 µM NCA for 6 h and analyzed by live cell imaging. Nuclei shown in blue, ER in red, bright-field (BF) shown to display coincidence of ER dye and vacuoles (scale bar 25 µm). C Data are presented in an XY graph as mean ± SD, n = 3. Statistical significance was analyzed by two-way ANOVA with Tukey’s posttest compared to mean of the corresponding DMSO control (ns not significant, *P < 0.05, **P < 0.01, ***P < 0.001).

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