Fig. 4

IPA alters mitochondrial dynamics and mtDNA amount in LX-2 cells. A Representative confocal images of live LX-2 cells with TGF-β1 (5 ng/ml) and IPA 1 mM in non-serum media for 24 h showing the mitochondrial network stained by Mitotracker™ Red CMXRos and blue nucleus with DAPI. All data were presented with at least 15 images per group. We acquired 10 Z-stack images from each sample type. Each Z-stack included 30 slices, with each slide having a thickness of 9.86 µm. Scale bar, 10 µm. B Representative objects identified by application of adaptative thresholding to the images (only mitochondria). Quantitative analysis and comparison of morphological mitochondrial network connectivity were performed on all cells in each group. C Frequency of proportion of mito shape. Values close to 0 mean spheres and close to 1 mean filamentous. D The mitochondria DNA amount (mtDNA) was measured as previously described in Material and Methods. E Mitotracker^™ Red CMXRos analyses were performed by flow cytometry (30.000 events) as previously described in Material and Methods. Data were shown as mean ± SD, n = 3 independent experiments. One-way ANOVA with Bonferroni´s post hoc test was used for statistical comparisons. *p < 0.05; **p < 0.01 ***p < 0.001; and ****p < 0.0001