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Fig. 2 | Journal of Experimental & Clinical Cancer Research

Fig. 2

From: Identification of distinct slow mode of reversible adaptation of pancreatic ductal adenocarcinoma to the prolonged acidic pH microenvironment

Fig. 2

Short- and long-term effects of extracellular acidity on PDAC mitochondrial network dynamics and bioenergetic reprogramming. a-b Representative N-SIM images (scale bar, 10 μm) show the time effects of extracellular acidosis on mitochondrial network dynamics in PDAC cells, which were processed by Imaris software for mitochondrial distribution and heterogeneity. The color scale from blue to red represents the different lengths of the mitochondrial tubules between branch points (0 to 9 μm). c TEM images of the time effects of acidic pHe stress on mitochondrial cristae ultrastructure. The lower panels are magnified areas derived from the respective upper panels as indicated by the boxed region. Scale bar: 2 μm. d-f The dynamic nature of cristae remodeling analyzed by ImageJ software with three column charts representing the number of total cristae, normal cristae, and distorted cristae per unit of mitochondrial cross-sectional areas as mean ± SD (n = 45 mitochondria). g Quantification of mitochondrial morphology from N-SIM images with three cells per condition randomly selected and combined to draw the full range of mitochondrial length distributions. h Representative blots of mitochondrial dynamics-related proteins under different control and acidic pHe stress conditions. i Western blots of OPA1 oligomerizations with VDAC as the loading control. j-o Cellular ROS, mitochondrial superoxide, and membrane potential were analyzed by confocal microscopy (j-l) and flow cytometry (m-o) using CellROX Green, MitoSOX Red, and TMRE Red probes, respectively. The relative fluorescence intensity of tumor cells in the absence (dotted line) or presence (solid line) of probes was shown in the histograms on the left. The corresponding values of fluorescence intensity were presented in the right column graphs as mean ± SD (n = 3). Scale bar, 50 μm. NS: p > 0.05 (not significant), *p < 0.05, ***p < 0.001

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