Journal: Translational Cancer Research

Article Title: Identification of a novel ion channel-related gene signature to predict prognosis and immune response of gastric cancer

doi: 10.21037/tcr-2025-1-2874

Figure Lengend Snippet: Knockdown of CNGB3 suppressed the intracellular Ca 2+ concentration and upregulated the expression of MHC-I in GC cells. (A,B) Intracellular Ca 2+ levels following CNGB3 gene knockdown. Confocal illustration of intracellular Ca 2+ imaging (A) and quantification (B). Scale bar: 100 μm (staining method: Fluo-4 AM staining; magnification: 100×). (C-E) Expression level of MHC-I molecules in cells after CNGB3 gene knockdown. **, P<0.01; ***, P<0.001. GC, gastric cancer; MHC-I, major histocompatibility complex class I.

Article Snippet: Cellular Ca 2+ levels were measured using a Fluo-4 AM Fluorescence Calcium Ion Detection Kit (Servicebio, Wuhan, China).

Techniques: Knockdown, Concentration Assay, Expressing, Imaging, Staining, Immunopeptidomics

Journal: Microorganisms

Article Title: Lsm1 Coordinates Mitochondrial Homeostasis, TORC1 Signaling, and Virulence in Candida albicans

doi: 10.3390/microorganisms14040771

Figure Lengend Snippet: Loss of Lsm1 induces mitochondrial dysfunction. ( A ) Localization of the PB marker GFP-Lsm1 with MitoTracker Red. BF, bright field; GFP, fluorescence showing the localization of GFP-Lsm1; MitoTracker Red, mitochondria staining; Merge, overlay of GFP-Lsm1 and MitoTracker Red signals. Arrows highlight GFP-Lsm1 puncta near mitochondria. Scale bar, 5 μm. ( B ) Fluorescence intensity plot profile analysis (line scan) corresponding to the dashed line in ( A ) using the Image J software (version 1.54f). The spatial overlap of green (GFP-Lsm1) and red (MitoTracker) peaks demonstrates co-localization of P-bodies on the mitochondrial surface. ( C ) Relative expression levels of genes involved in mitochondrial respiratory chain complexes in WT and lsm1 Δ/Δ cells, shown as Log 2 fold change and the percentage of differentially expressed genes. Significant downregulation ( p < 0.05, blue). Genes involved in the mitochondrial respiratory chain with significant transcriptional changes are shown. ( D ) Mitochondrial membrane potential (ΔΨm) was quantified by flow cytometric analysis using the JC-1 in WT, lsm1 Δ/Δ and complemented ( lsm1 Δ/Δ+ LSM1 ) strains. Depolarized mitochondria were detected as JC-1 monomers (green fluorescence, Ex 488 nm/Em 530 nm), whereas healthy, hyperpolarized mitochondria were characterized by JC-1 J-aggregates (red fluorescence, Ex 488 nm/Em 590 nm). The red lines indicate the flow cytometry gates used to distinguish the populations of JC-1 aggregates (upper region) and JC-1 monomers (lower region). ( E ) Intracellular reactive oxygen species (ROS) levels were measured using the fluorescent probe DCFH-DA in WT, lsm1 Δ/Δ and lsm1 Δ/Δ+ LSM1 strains. ( F ) Cytosolic Ca 2+ levels were determined using the calcium-sensitive fluorescent dye Fluo-4 AM. ( G ) Mitochondrial Ca 2+ levels were assessed using the mitochondria-targeted Ca 2+ indicator Rhod-2 AM. Statistical significance is indicated as follows: ***, p < 0.001; ns, not significant.

Article Snippet: Intracellular calcium levels were determined following the same procedure, using the calcium-sensitive dye Fluo-4 AM (2 μM, MCE, USA) for cytosolic Ca 2+ and the mitochondria-targeted indicator Rhod-2 AM (10 μM, MCE, USA) for mitochondrial Ca 2+ levels.

Techniques: Marker, Fluorescence, Staining, Software, Expressing, Membrane, Flow Cytometry