Journal: American Journal of Cancer Research

Article Title: Mitochondrial calcium uniporter as a biomarker in lung adenocarcinoma: regulation of the immune microenvironment, apoptosis, cell cycle and mitophagy

doi: 10.62347/PGQK3358

Figure Lengend Snippet: MCU regulates mitophagy and mitochondrial function. A. GSEA analysis showed that MCU-associated genes were significantly enriched in the “autophagy of mitochondrion” and “positive regulation of autophagy of mitochondrion” pathways; B. Western blot analysis was performed to detect the expression of mitophagy-related proteins, including PINK1, PRKN, and LC3B, in H1975 and PC9 cells; C, D. Immunofluorescence analysis was used to examine the colocalization of mitochondria and LC3B in H1975 and PC9 cells; E, F. JC-1 staining combined with flow cytometry was performed to assess changes in mitochondrial membrane potential in H1975 and PC9 cells; G, H. Flow cytometry was used to detect intracellular ROS levels in H1975 and PC9 cells. Data are presented as the mean ± SD. Statistical analysis was performed using two-sided t-tests. **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: JC-1 assay kit (MedChemExpress, New Jersey, USA) is to be used for the mitochondrial membrane potential experiment.

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Flow Cytometry, Membrane

Journal: Genes & Diseases

Article Title: PDK1 elevation was induced by epigenetic modifications of KDM3A and METTL16 to mediate TKI resistance and cancer development

doi: 10.1016/j.gendis.2025.101947

Figure Lengend Snippet: PDK1 inhibitor JX06 and gefitinib synergistically induced cell apoptosis in gefitinib-resistant lung cancer cells. (A) The protein expression levels of PDK were reduced upon the treatment of PDK1 inhibitor JX06 in PC-9 and PC-9/G cells. (B) B2B and PC-9/G cells were treated with different concentrations of JX06 for 48 h. The cell viabilities were determined by CCK-8. (C – E) The synergy effect between JX06 and gefitinib was determined and analyzed with CompuSyn software. (F) The apoptosis rates were analyzed with flow cytometry after the combined treatment of gefitinib and JX06. (G) The TUNEL assay was performed with the indicated treatment in PC-9/G cells. (H) The cells treated as described were stained with the JC-1 probe and detected using a fluorescence microscope. Red fluorescence indicates the aggregation form of JC-1, showing increased mitochondrial membrane potential (ΔΨm). Green fluorescence indicates the monomeric form of JC-1, which indicates reduced mitochondrial membrane potential (ΔΨm). Data were statistically analyzed with Student’s t -test, and values were shown as mean ± standard deviation. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: After different treatments for 48 h, the mitochondrial membrane potential of cells was detected with the enhanced mitochondrial membrane potential assay kit using JC-1 probe (Beyotime, Jiangsu, China), and the fluorescence was analyzed with a fluorescence microscope.

Techniques: Expressing, CCK-8 Assay, Software, Flow Cytometry, TUNEL Assay, Staining, Fluorescence, Microscopy, Membrane, Standard Deviation

Journal: Materials Today Bio

Article Title: Ultrasmall Prussian blue–integrated cryogel for enhanced ROS scavenging and immunomodulation via cGAS–STING inhibition in wound healing

doi: 10.1016/j.mtbio.2026.103056

Figure Lengend Snippet: Schematic representation of the synthesis process and therapeutic properties of Cryogel@USPB applied in wounds. (A) Fabrication process of Cryogel@USPB. (B) Cryogel@USPB promotes acute/chronic wounds through regulating macrophage polarization and restraining mitochondrial dysfunction via the cGAS-STING pathway.

Article Snippet: The Catalase Assay Kit and Mitochondrial Membrane Potential Assay Kit (JC-1) were from Beyotime Biotechnology Inc. (Shanghai, China).

Techniques:

Journal: Materials Today Bio

Article Title: Ultrasmall Prussian blue–integrated cryogel for enhanced ROS scavenging and immunomodulation via cGAS–STING inhibition in wound healing

doi: 10.1016/j.mtbio.2026.103056

Figure Lengend Snippet: The biocompatibility and antioxidant capability assessment of Cryogel@USPB. (A) Cell viability of NIH 3T3 and Raw264.7 cells after co-incubation with Cryogel@USPB at various concentrations (n = 3). (B) Representative fluorescent staining images of live (green)/dead (red) cells of Raw264.7 cells following co-incubation with Cryogel@USPB. (C) Determination of H 2 O 2 depletion capability of Cryogel@USPB (n = 3). (D) SOD-like activity and (E) POD-like activity of Cryogel@USPB (n = 3). (F) ·OH depletion capability and (G) •O 2 − depletion activity using EPR. (H) Representative fluorescent staining images of intracellular ROS in Raw264.7 cells following the treatment of H 2 O 2 and Cryogel@USPB. (I) TEM images of the mitochondria in RAW264.7 cells. (J) Fluorescent images of mitochondrial membrane potential in cells following different treatments. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The Catalase Assay Kit and Mitochondrial Membrane Potential Assay Kit (JC-1) were from Beyotime Biotechnology Inc. (Shanghai, China).

Techniques: Incubation, Staining, Activity Assay, Membrane

Journal: Human Mutation

Article Title: PLSCR3 Deficiency Triggers mtDNA‐Driven cGAS‐STING Activation to Potentiate Antitumor Immunity in Colorectal Cancer

doi: 10.1155/humu/8545428

Figure Lengend Snippet: PLSCR3 deficiency disrupts mitochondrial homeostasis in CRC cells. (A) Western blotting verified the reduction of PLSCR3 protein levels in HT29 cells. (B) RT‐qPCR analysis verified reduction of PLSCR3 mRNA levels normalized to GAPDH compared with negative control (NC) cells ( p < 0.0001, n = 3). (C) Representative JC‐1 staining (scale bar: 20 μ m) images showing red fluorescence (polarized mitochondria) and green fluorescence (depolarized mitochondria). (D) Quantification of the JC‐1 red/green fluorescence intensity ratio. Fluorescence intensity was quantified using ImageJ software as described in the section. (E) Real‐time oxygen consumption rate (OCR) profiles under sequential treatment with oligomycin, FCCP, and rotenone/Antimycin A. (F) Quantification of cytosolic mtDNA by qPCR using the D‐loop ratio in PLSCR3‐knockdown and control cells. Data are presented as mean ± SD from three independent experiments. Statistical significance was determined using an unpaired two‐tailed Student′s t ‐test for two‐group comparisons.

Article Snippet: Cells were incubated with JC‐1 dye (MedChemExpress, Cat. No. HY‐K0601) at 37°C in the dark, enabling accumulation in mitochondria; high membrane potential promoted J‐aggregate formation (red fluorescence), while depolarized mitochondria retained JC‐1 monomers (green fluorescence).

Techniques: Western Blot, Quantitative RT-PCR, Negative Control, Staining, Fluorescence, Software, Knockdown, Control, Two Tailed Test