Fig. 4

Decrease of mitochondrial mass and intracellular ATP is accompanied with morphological changes of mitochondria in tumor cell lines overexpressing TNFα. a Measurement of intracellular ATP (Luminescent Cell Viability Assay, Promega) in exponentially growing cells; means of pentaplicates + SDs, unpaired t-test was used to statistical analysis. b Staining of mitochondrial mass with MitoTracker DeepRedFM (25 nM) (left) and detection of mitochondrial activity with MitoTracker OrangeCMT (250 nM) (right) in cells cultivated for 3–4 days; flow cytometry, values are expressed as the means of triplicates + SDs, unpaired t-test was used to statistical analysis. c Fluorescence microscopy of mitochondrial morphology in adherent cells, red – mitochondria stained with Mitotracker OrangeCMT, blue – nuclei/DAPI; white arrows – condensed/shortened mitochondria in hTNFα overexpressing cells; magnification 630x