fluorescent ca 2 indicator fluo 3 am  (MedChemExpress)

Journal: Science Advances

Article Title: Adaptable thermoresponsive polymer for long-term electrical coupling in plant electrophysiology monitoring

doi: 10.1126/sciadv.ady1400

Figure Lengend Snippet: ( A ) Procedure of drought stress and recovery experiment and photographs of plants under testing. Predrought is defined as 1 day before plants display wilting symptoms. ( B ) Amplitude of light-induced potential change for plants under 9 days of escalating drought (red) benchmarked against well-watered control plants (gray), showing an increase followed by decrease upon wilting. a.u., arbitrary units. ( C ) Light-induced potential change before and after inhibition of Ca 2+ channels in the same leaf. N = 3. ( D ) Light-induced potential change before and after inhibition of ROS production in the same leaf. N = 3. ( E ) Confocal images of intracellular Ca 2+ (top) and apoplastic ROS (bottom) during drought simulation, with quantification of the mean fluorescence intensity per region of interest (ROI) shown on the right. BF, bright field. Fluo-3 AM and 2′,7′-dichlorodihydrofluorescein diacetate (H 2 DCFDA): dyes used for visualizing Ca 2+ and ROS, respectively. Scale bar, 50 μm. CK, control plants; DR, drought-stressed plants. ( F ) Plant EP signals under the light and dark conditions with starting potential aligned for all samples on each day. N = 6. ( G ) Mean potential calculated from signals in (F) 1 hour after light on or light off, showing opposing trends with time under light and dark conditions. Dark mean potential changes before visual symptom onset on day 7. Solid lines in (C), (D), and (F) are mean values, and shadows represent SD. Box plots in (B), (E), and (G) display first quartile, median, and third quartile. Whiskers are drawn at 1.5 interquartile range distance. Means are denoted by empty squares. * P < 0.05; *** P < 0.001; **** P < 0.0001; n.s., not significant. Two sample t test.

Article Snippet: Intracellular Ca 2+ ions were detected using the fluorescent Ca 2+ indicator Fluo-3 AM (MedChemExpress, HY-D0716).

Techniques: Control, Inhibition, Fluorescence

Journal: Heliyon

Article Title: VDAC1, as a downstream molecule of MLKL, participates in OGD/R-induced necroptosis by inducing mitochondrial damage

doi: 10.1016/j.heliyon.2023.e23426

Figure Lengend Snippet: MLKL is involved in OGD-induced mitochondrial damage processes through the oligomerization of VDAC1. (A) Fluo-3 AM tag was used to detect the changes in the concentration of calcium ions in NIH3T3 cells after OGD injury, and Fluo-3 AM combined with calcium ions emits green fluorescence; Scale bars: 200 μm. (B) The intracellular concentration of the calcium ion detection kit was used to assess changes in intracellular calcium ion concentrations in NIH3T3 cells after OGD injury. *** p < 0.001 vs. CTL group; # p < 0.05 vs. OGD3 hr R 4 h group; ## p < 0.01 vs . OGD3 hr R 4 h group. (C) Calcium ion fluorescence intensity was detected by flow cytometry after Fluo-3 AM treatment. (D) Changes in the VDAC1 oligomerization form in whole cell extraction treated with EGS (ethylene glycol bis (succinimidyl succinate)) after OGD injury. (E) Statistical analysis of the ratio of oligomerization VDAC1 to total VDAC1. *** p < 0.001 vs. CTL group; **** p < 0.001 vs. CTL group; ## p < 0.01 vs. OGD3 hr R 4 h group; ### p < 0.001 vs. OGD3 hr R 4 h group. (F) Statistical analysis of the VDAC1 oligomerization/beta Actin in total cell extraction treated with EGS after OGD injury. ** p < 0.01 vs. CTL group; *** p < 0.001 vs. CTL group; # p < 0.05 vs. OGD3 hr R 4 h group. (G) JC-1 was used to detect mitochondrial dysfunction, CTL mitochondrial membrane potential (red), and depolarized membrane potentials (green).

Article Snippet: Intracellular Ca 2+ levels were evaluated using the fluorescent Ca 2+ indicator fluo-3-acetoxymethylester AM (Fluo-3 AM, Beyotime, Cat#S1056) [ , ].

Techniques: Concentration Assay, Fluorescence, Flow Cytometry, Extraction, Membrane

Journal: Journal of Translational Medicine

Article Title: Estrogen antagonizes ASIC1a-induced chondrocyte mitochondrial stress in rheumatoid arthritis

doi: 10.1186/s12967-022-03781-1

Figure Lengend Snippet: ASIC1a upregulated mitochondrial stress by mediating Ca 2+ influx. A The effect of ASIC1a on Ca 2+ in chondrocytes at a pH of 6.0, as detected by calcium imaging. The scale bars are 20 μm. B – D Hsp10 , Lonp1 , and ClpP mRNA levels in acid-mediated chondrocytes after EGTA pretreatment quantified by real-time q-PCR. E – G HSP10, LONP1, and ClpP expression in acid-mediated chondrocytes after EGTA pretreatment quantified immunofluorescence. The scale bars are 20 μm. H Measurements of ROS released by mitochondria in chondrocytes after pre-treatment with EGTA. I Representative confocal microscopy images of chondrocytes stained with JC-1 after pre-treatment with E2 and EGTA. Red fluorescence indicates normal ΔΨm with JC-1 aggregates in the mitochondria, and green fluorescence reflects the JC-1 monomer, indicative of ΔΨm loss. The scale bars are 20 μm. Data are presented as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Cytosolic Ca 2+ levels in isolated rat articular chondrocytes were measured using the fluorescent Ca 2+ indicator Fluo-3 AM (Dojindo Laboratories).

Techniques: Imaging, Expressing, Immunofluorescence, Confocal Microscopy, Staining, Fluorescence

Journal: Journal of Translational Medicine

Article Title: Estrogen antagonizes ASIC1a-induced chondrocyte mitochondrial stress in rheumatoid arthritis

doi: 10.1186/s12967-022-03781-1

Figure Lengend Snippet: Estrogen decreased ASIC1a activity in articular chondrocytes. A The expression of ASIC1a decreased in a time-dependent manner following stimulation with 1000 nmol/mL estradiol for 0–48 h in chondrocytes. B Immunofluorescence analysis of ASIC1a expression in chondrocytes treated with PcTx-1, E2, and EGTA. The scale bars are 20 μm. C Calcium imaging detected the effect of ASIC1a on Ca 2+ in chondrocytes treated with E2, PcTx-1. The scale bars are 20 μm. D Calcium imaging revealed ASIC1a-mediated Ca 2+ influx with E2, PcTx-1. Data are presented as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Cytosolic Ca 2+ levels in isolated rat articular chondrocytes were measured using the fluorescent Ca 2+ indicator Fluo-3 AM (Dojindo Laboratories).

Techniques: Activity Assay, Expressing, Immunofluorescence, Imaging

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Polydatin Attenuates OGD/R-Induced Neuronal Injury and Spinal Cord Ischemia/Reperfusion Injury by Protecting Mitochondrial Function via Nrf2/ARE Signaling Pathway

doi: 10.1155/2021/6687212

Figure Lengend Snippet: PD restores mitochondrial functions in OGD/R-injured neurons. (a) [Ca 2+ ] i levels in cells exposed to normoxia or hypoxia-reperfusion in the indicated groups. (b) Normalized relative fluorescence units (NRFU) of calcein indicating mPTP opening in the differentially treated cells. (c) The ratios of polymeric (red) and monomeric (green) forms of JC-1 corresponding to the MMP in the indicated groups. Scale bars = 100 μ m. (d) MitoSOX fluorescence intensity indicative of ROS levels. Data are presented as fold change over the Control group. Scale bars = 10 μ m. (e) Neuronal ATP release (nmol/mg protein) as measured by a luciferase-based assay. ∗ P < 0.05 vs. Control; # P < 0.05 vs. OGD; ΔP < 0.05 vs. OGD + PD (15 μ M).

Article Snippet: Free intracellular Ca 2+ ([Ca 2+ ] i ) levels were measured by staining with the fluorescence Ca 2+ indicator Fluo-3/AM (Beyotime).

Techniques: Fluorescence, Control, Luciferase

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Polydatin Attenuates OGD/R-Induced Neuronal Injury and Spinal Cord Ischemia/Reperfusion Injury by Protecting Mitochondrial Function via Nrf2/ARE Signaling Pathway

doi: 10.1155/2021/6687212

Figure Lengend Snippet: PD-mediated mito-protective depends on the Nrf2/ARE pathway. (a) [Ca 2+ ] i levels in cells exposed to normoxia or hypoxia-reperfusion in the indicated groups. (b) NRFU of calcein indicating mPTP opening in the differentially treated cells. (c) The ratios of polymeric (red) and monomeric (green) forms of JC-1 corresponding to the MMP in the indicated groups. (d) MitoSOX fluorescence intensity indicative of ROS levels. Data are presented as fold change over the Control group. Scale bars = 10 μ m. (e) Neuronal ATP release (nmol/mg protein) as measured by a luciferase-based assay. ∗ P < 0.05 vs. Control; # P < 0.05 vs. OGD; ΔP < 0.05 vs. OGD + PD.

Article Snippet: Free intracellular Ca 2+ ([Ca 2+ ] i ) levels were measured by staining with the fluorescence Ca 2+ indicator Fluo-3/AM (Beyotime).

Techniques: Fluorescence, Control, Luciferase

Journal: Cell Death & Disease

Article Title: Gp130-mediated STAT3 activation by S -propargyl-cysteine, an endogenous hydrogen sulfide initiator, prevents doxorubicin-induced cardiotoxicity

doi: 10.1038/cddis.2016.209

Figure Lengend Snippet: Suppression of gp130/STAT3 reduces the inhibitory effect of SPRC on Dox-induced [Ca 2+ ] i overload. Cardiomyocytes were pretreated with/without SC144 before SPRC incubation, followed by Dox (1 μ M) stimulation for 24 h. The intracellular [Ca 2+ ] i concentration was ( a ) quantified by flow cytometry and ( b ) visualized by fluorescence microscopy using fluo-3/AM probe, a fluorescent Ca 2+ -indicator dye. Scale bars, 50 μ m. ( c ) Representative western blot analysis of SERCA2. Values are presented as mean±S.D. from n =3 replicates. # P <0.05, ### P <0.001 compared with the control group; * P <0.05, *** P <0.001 compared with the Dox group. NS, nonsignificant ( P >0.05)

Article Snippet: The level of intracellular [Ca 2+ ] i was measured using fluo-3/AM (Dojindo Lab), a fluorescent Ca 2+ -indicator probe.

Techniques: Incubation, Concentration Assay, Flow Cytometry, Fluorescence, Microscopy, Western Blot, Control

Journal: Cell Death & Disease

Article Title: Gp130-mediated STAT3 activation by S -propargyl-cysteine, an endogenous hydrogen sulfide initiator, prevents doxorubicin-induced cardiotoxicity

doi: 10.1038/cddis.2016.209

Figure Lengend Snippet: Schematic illustration of proposed mechanism of gp130/STAT3 signaling modulation by SPRC. SPRC activates STAT3 though gp130 in Dox-induced cardiomyocytes and hearts. SPRC attenuates Dox-induced cardiotoxicity via a mechanism involving the promotion of gp130-mediated STAT3 activity, leading to activation of STAT3-regulated cardioprotective molecules expression (e.g., MCL-1, Bcl-2, Bcl-X L , Survivin and MnSOD), mitigation of mitochondrial dysfunction, and suppression of ROS generation and [Ca 2+ ] i accumulation

Article Snippet: The level of intracellular [Ca 2+ ] i was measured using fluo-3/AM (Dojindo Lab), a fluorescent Ca 2+ -indicator probe.

Techniques: Activity Assay, Activation Assay, Expressing