parp  (Cell Signaling Technology Inc)

Journal: Redox Biology

Article Title: Mitochondrial permeability transition pore desensitization by a novel dispiranic derivative prevents cardiac reperfusion injury

doi: 10.1016/j.redox.2026.104097

Figure Lengend Snippet: Cytoprotective effects following H/R. (A) Calcein−cobalt assay in AC16 cells. The kinetics of PTP opening were evaluated at the moment of the reoxygenation in the presence of vehicle, 11d or 12c compounds. The raw values were expressed as a percentage of the vehicle. Each value is the mean of at least 10 cells from 3 biological and 3 technical replicates. (B) Quantification of AC16 cells positive to Propidium Iodide (PI + ) staining following H/R. Each value is the mean of 3 biological and 4 technical replicates. (C) Calcein-cobalt assay in differentiated HCM cells. Data has been collected at reperfusion time in the presence of vehicle or 11d compound. (D) Immunoblot detection of the main markers of apoptosis and necrosis such as Cleaved PARP, Cleaved Caspase 3 and Cleaved RIP. This is representative of 3 biological replicates. (E) Quantification of cell viability according to viable cells upon H/R staining with crystal violet. This is representative of at least 3 biological and 3 technical replicates. (F) Quantification of the MitoSox intensity of cells after H/R under the same experimental conditions. Each value is the mean of at least 15 cells from 3 technical and 3 biological replicates. (G) Mitochondrial calcium uptake in AC16 cells by using a mitochondrially targeted aequorin probe (mtAeq); [Ca 2+ ] is detected at the time of reoxygenation and after addition of 100 μM His and 100 μM Bk. Each value is the mean of at least of 3 biological and 3 technical replicates. Histograms of statistical and representative kinetics data are reported. One-way ANOVA was applied for statistical analysis for all graphs reported in the figure; (∗∗∗∗) p value < 0.0001; (∗∗∗) p value < 0.001; (∗∗) p value < 0.01; (∗) p value < 0.05.

Article Snippet: After electrophoretic separation, proteins were transferred onto nitrocellulose membranes that were incubated overnight with the following primary antibodies: cleaved PARP (Cell signaling, 9541, 1:1000), Caspase 3 (Cell signaling, 9662, 1:1000), RIP (Cell signaling, 3493, 1:1000), β-Actin (Merck, A1978, 1:5000), ATP5A (Abcam, ab14748, 1:5000).

Techniques: Cobalt Assay, Staining, Western Blot

Journal: Oncology Reports

Article Title: Sphingosine-1-phosphate receptor 1 enhances olfactory receptor 51E1-mediated inhibition of proliferation via Src/JNK signaling in prostate cancer cells

doi: 10.3892/or.2026.9103

Figure Lengend Snippet: S1PR1 enhances OR51E1-mediated cytotoxicity and apoptosis in LNCaP cells. (A) LNCaP cells were transfected with an empty vector, S1PR1, or DRD2. After 48 h, cells were treated with NA (0.5 mM) for an additional 48 h, and cell viability was measured using the Cell Counting Kit-8 assay. (B) LNCaP cells were transduced with lentivirus encoding S1PR1 and selected with puromycin (1 µg/ml). Cell viability of parental LNCaP and LNCaP-S1PR1 cells was measured 48 h after treatment with increasing concentrations of NA (0.1–1 mM). (C) Apoptosis was assessed in LNCaP cells treated with NA for 48 h using annexin V/PI staining. Apoptotic cells were quantified as the percentage of annexin V-positive cells (early apoptosis) and annexin V/PI double-positive cells (late apoptosis) relative to the total cell population. (D) Caspase-3 activation was measured by flow cytometry following NA treatment. (E) PARP1 cleavage was analyzed by western blotting in NA-treated cells. Band intensities were quantified using ImageJ software. Data represent the mean ± SEM of at least three independent experiments. Statistical significance was determined using an unpaired Student's t-test. *P<0.05, **P<0.01 and ***P<0.001. S1PR1, sphingosine-1-phosphate receptor 1; OR51E1, olfactory receptor 51E1; DRD2, dopamine receptor D2; NA, nonanoic acid; PI, propidium iodide; ns, not significant.

Article Snippet: Anti-S1PR1 (cat. no. 55133-1-AP) and anti-PARP1 (cat. no. 51-6639GR) antibodies were purchased from Proteintech Group, Inc. and BD Biosciences, respectively.

Techniques: Transfection, Plasmid Preparation, Cell Counting, Transduction, Staining, Activation Assay, Flow Cytometry, Western Blot, Software, Olfactory