Journal: Oxidative Medicine and Cellular Longevity

Article Title: Slowly Repaired Bulky DNA Damages Modulate Cellular Redox Environment Leading to Premature Senescence

doi: 10.1155/2020/5367102

Figure Lengend Snippet: PUVA-induced mitochondrial mass increase and the suppression by ribose-5-phosphate. (a–c) The mitochondrial mass in control (a) and PUVA-treated (at week 3 and week 6) fibroblasts (b, c) determined by MitoTracker Red; (d) determination of mitochondrial membrane potential ( ΔΨ ) by a polarization-sensitive fluorescent dye JC-9 based on the ratio between two ΔΨ -driven interchangeable forms, monomer (J-monomer, green) and dimer (J-aggregate, red); (e, f) the cell morphology after PUVA (at week 3) without (e) and with ribose-5-phosphate (5 mM, f).

Article Snippet: Mitochondrial mass was determined by staining living cells with 25 nM of the fluorescent dye MitoTracker Red CMXRos (MoBiTec, Goettingen) in DMEM for 30 min at 37°C, then photograph using a 568 nm line of fluorescence excitation with a laser scanning confocal microscope.

Techniques: Control, Membrane

Journal: Microbiology Spectrum

Article Title: The mitochondrial protein Bcs1A regulates antifungal drug tolerance by affecting efflux pump expression in the filamentous pathogenic fungus Aspergillus fumigatus

doi: 10.1128/spectrum.01172-24

Figure Lengend Snippet: Mitochondria-localized Bcs1A is required for hyphal growth in A. fumigatus . ( A ) Bioinformatic analysis of selected Bcs1 homologs from eukaryotes. The phylogenetic tree was constructed using MEGA 7 software. Domains were analyzed using SMART ( http://smart.embl-heidelberg.de/ ). ( B ) Subcellular localization of green fluorescent protein (GFP)-tagged Bcs1A. MitoTracker was used to visualize mitochondria. Scale bar, 10 µm. Colony phenotypes ( C ) and diameters ( D ) of the wild-type (WT), Δ bcs1A , and bcs1A C strains grown in minimal medium (MM) and yeast extract-glucose (YG) media at 37°C for 48 h. ns, not significant; ***, P < 0.001.

Article Snippet: To visualize the subcellular distribution of Bcs1A, the GFP-labeled strain was incubated in 500 μL of liquid MM on glass coverslips for a duration of 12 h. Samples were fixed in 4% formaldehyde for 30 min and then incubated in the dark with the mitochondrial dye MitoTracker Red CMXRos (Beyotime) for 5 min used at a final concentration of 5 nM.

Techniques: Construct, Software

Journal: Acta Pharmaceutica Sinica. B

Article Title: Astaxanthine attenuates cisplatin ototoxicity in vitro and protects against cisplatin-induced hearing loss in vivo

doi: 10.1016/j.apsb.2021.07.002

Figure Lengend Snippet: Effects of astaxanthine on maintaining mitochondrial function in HEI-OC1 cell line and IHCs. (A) and (B) Representative MTRC (red) and DAPI (blue) stained confocal images of HEI-OC1 cells from the control, CDDP, AST, and AST/CDDP groups. The ImageJ was used for the analysis of quantitative changes in the mean MTRC fluorescence intensity. The data are presented as mean ± SD, ∗∗∗∗ P < 0.0001 compared with control (blue) and CDDP (black) by one-way ANOVA with Bonferroni correction, n = 6 for each group in (B). Scale bar = 10 μm. (C) and (D) Representative MTRC (magenta) and DAPI (blue) stained confocal images of IHCs in middle regions of postnatal 3-day C57BL/6 mouse cochleae from different treatment groups. The Ortho section shows mitochondrial staining of IHCs and the ROI fluorescence from a single IHC in each group was identified under the same microscope settings (C). The ImageJ was used for the analysis of quantitative changes in the mean MTRC fluorescence intensity (D). The data are presented as mean ± SD, ∗∗∗ P < 0.001 compared with control (blue) and CDDP (black) by one-way ANOVA with Bonferroni correction, n = 3 for each group in (D). Scale bar = 20 μm. ns, not significant; AST, astaxanthine; CDDP, cisplatin; IHC, inner hair cell; MTRC, MitoTracker Red CMXRos; ROI, region of interest.

Article Snippet: The fluorescent dye MitoTracker Red CMXRos (MTRC; Beyotime) was used for monitoring MMP.

Techniques: Staining, Control, Fluorescence, Microscopy