mitotracker red cmxros  (Beijing Solarbio Science)

Journal: International Dental Journal

Article Title: Inhibition of Dynamin-Related Protein 1-Dependent Mitochondrial Fission Ameliorates Apical Periodontitis by Attenuating NLRP3 Inflammasome-Mediated M1 Macrophage Polarisation

doi: 10.1016/j.identj.2025.100853

Figure Lengend Snippet: Pg-LPS induces abnormal mitochondrial division, activation of NLRP3 inflammatory pathway and M1 polarisation in macrophages. (A) Representative western-blotting images and quantification in different groups. (B) Representative images of ROS staining and quantification of intensity units in each group. Scale bar = 50 μm. (C) Representative images of JC-1 staining in different groups and quantification of red and green fluorescence intensity. Scale bar = 50 μm. (D) Quantification data of ATP level in macrophages. (E) Representative immunofluorescence images identified the colocalisation of p-Drp1 (Ser616) /TOMM20. Plots of pixel intensity of p-Drp1 (Ser616) with TOMM20. Scale bar = 10 μm. (F) Representative images of MitoTracker Red stain immunofluorescence (white boxes indicate higher magnification area of corresponding lower panels). Quantification of mitochondrial number and length. n = 25 Scale bar = 10 μm. (G) Mitochondrial morphology was visualised by transmission electron microscopy. Scale bar = 1 μm. All data were analysed by one-way ANOVA and presented as mean ± SD (n = 3). * P < .05, ** P < .01, *** P < .001.

Article Snippet: After the different stimulation, cells were processed according to staining requirements: for MitoTracker staining, cells were plated on the confocal petri dish and incubated with MitoTracker Red CMXRos (Solarbio, China) for 0.5 hours at 37°C.

Techniques: Activation Assay, Western Blot, Staining, Fluorescence, Immunofluorescence, Transmission Assay, Electron Microscopy

Journal: Journal of Neuroinflammation

Article Title: Activated TBK1 promotes ACSL1-mediated microglia lipid droplet accumulation and neuroinflammation in Parkinson’s disease

doi: 10.1186/s12974-025-03517-0

Figure Lengend Snippet: ACSL1 knockdown inhibits lipid droplet formation. a-b , Reactome and KEGG enrichment was used to visualize the metabolic pathways of ACSL1 high microglia. c-d , Transfected BV2 cells were treated with PBS or LPS (1 µg/ml) for 18 h. Representative micrographs of BODIPY-stained BV2 cells. (c) Quantification of the BODIPY fluorescence per cell (d). e , Schematic representation of the structural composition of an LD. Coloured objects represent LD surface-bound proteins localized to the phospholipid monolayer. Triacylglycerols and sterol esters are found in the neutral lipid core. f , Representative confocal images of transfected BV-2 cells after treatment with LPS and coadministration of chloroquine (20 µM) for 12 h. Scale bar, 20 μm. g , The expression of p62 and LC3B was assessed by Western blot in BV-2 cells transfected with NC or si-ACSL1 after treatment with LPS and cotreated with chloroquine. h-i , The relative mRNA expression of lipolysis markers (h) and lipid uptake markers (i) in BV-2 cells transfected with NC or si-ACSL1 after treatment with LPS or PBS. j , Colocalization of BODIPY (green) and ACSL1 (red) in BV-2 cells. k , The expression levels of ACSL1 in the mitochondrial and cytosolic fractions of BV-2 cells were assessed by Western blotting. Tomm20 was used as a mitochondrial fraction marker. l , The expression levels of ACSL1 in the ER fractions of BV-2 cells were assessed by Western blotting. Calreticulin was used as an ER fraction marker. m , The colocalization of ACSL1 (green) and MitoTracker (red) in BV-2 cells was analysed by confocal microscopy. Quantification of fluorescence colocalization was performed with ImageJ. All data are shown as mean ± SEM using one-way ANOVA. Multiple comparison corrections were applied via Dunnett’s multiple comparisons test. *** P < 0.001; ** P < 0.01; * P < 0.05; ns, not significant

Article Snippet: The mitochondria of the cells were labelled with MitoTracker Red CMXRos (Beyotime).

Techniques: Knockdown, Transfection, Staining, Fluorescence, Expressing, Western Blot, Marker, Confocal Microscopy, Comparison