Journal: Frontiers in Immunology

Article Title: P-glycoprotein expression skews mitochondrial dye measurements in T cells

doi: 10.3389/fimmu.2025.1560104

Figure Lengend Snippet: P-glycoprotein (P-gp) expression affects mitochondrial dye measurements in T cells. (a, b) MTG signals from splenic naïve (CD44 - CD62L + ) and memory (CD44 + CD62L + ) CD8 T cells stained with or without PSC833 (1uM). (c) Protein expression level of P-glycoprotein based on Variance Stabilization Normalization (VSN) Normalized Intensities from proteomic analysis. (d, e) MTG and (f, g) TMRE signals from thymic CD4 T (CD8a - CD1d-aGC-tetramer - CD4 + TCRβ hi ) and iNKT (CD8a - CD1d-aGC-tetramer + TCRb int ) cells stained with or without PSC833 as in (a) . (h, i) MTG and (j, k) TMRE signals from PBMC CD4 T (CD8a - CD1d-aGC-tetramer - CD4 + TCRb hi ) and iNKT (CD8a - CD1d-aGC-tetramer + TCRb int ) cells stained with or without PSC833 as in (a) . The data shown in (a, b, d-k) are one representative experiment out of three independent experiments. The data shown in c are from samples collected from two sorting experiments and proteomes analyzed together. ns, non-significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, paired Student’s t test.

Article Snippet: PSC833 was from Santa Cruz Biotechnology.

Techniques: Expressing, Staining

Journal: Biomedical Reports

Article Title: Scutellarein enhances cisplatin‑induced apoptotic effects by suppressing the PI3K/AKT‑MDR1 pathway in human NPC/HK1 nasopharyngeal carcinoma cells

doi: 10.3892/br.2025.1938

Figure Lengend Snippet: Scutellarein enhances cisplatin-induced inhibition of cell viability and release of cytokeratin 18 fragments by inhibiting the PI3K/AKT-MDR1 pathway in NPC/HK1 cells. (A) NPC/HK1 cells were treated without (Ctrl) or with 12.5 µM scutellarein, 4.15 µM cisplatin, or their combination for 48 h. Protein expression of p-AKT, AKT, MDR1, and GAPDH was examined using an immunoblotting assay. (B) NPC/HK1 cells were treated without (Ctrl) or with 4.15 µM cisplatin, or 4.15 µM cisplatin plus 10 µM LY294002 for 48 h. Upper panel, protein expression of p-AKT, AKT, MDR1, and GAPDH was examined using an immunoblotting assay. Lower panel, cytokeratin 18 fragment levels in the cell culture supernatant were measured by ELISA. (C) NPC/HK1 cells were treated without (Ctrl) or with 4.15 µM cisplatin, or 4.15 µM cisplatin plus 10 µM PSC833 for 48 h. Upper panel, protein expression of MDR1 was examined using an immunoblotting assay. Middle panel, cell viability was assessed using the MTT assay. Lower panel, cytokeratin 18 fragment levels in the cell culture supernatant were measured by ELISA. Statistical significance was indicated as follows: * P<0.05 vs. Ctrl; and # P<0.05 vs. Cis alone; n=3. p-, phosphorylated; MDR1, multidrug resistance protein 1; Ctrl, control; Scu, scutellarein; Cis, cisplatin; LY, LY294002.

Article Snippet: Scutellarein, cisplatin, dimethyl sulfoxide (DMSO), 3-methyladenine (3-MA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), PSC833, and LY294002 were obtained from MilliporeSigma.

Techniques: Inhibition, Expressing, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, MTT Assay, Control

Journal: Cancers

Article Title: A Role for iNOS in Erastin Mediated Reduction of P-Glycoprotein Transport Activity

doi: 10.3390/cancers16091733

Figure Lengend Snippet: Changes in P-glycoprotein (P-gp) transport activity in brain capillaries ( N = 15–20 per data point) from six rats treated with erastin. ( A ) Representative confocal fluorescent and DIC images of brain capillaries after a 60-min incubation with 2.0 μM NBD-CSA; note the high luminal fluorescence in the control capillary and ( B ) decreased luminal fluorescence in capillaries exposed to 10.0 μM PSC833 (Scale bars, 10 μm). Representative confocal images of the luminal fluorescence (LF) from ( C ) male and female brain capillaries associated with P-glycoprotein (P-gp) transport activity at increasing doses of erastin in brain capillaries of SD rats. Bar graphs of P-glycoprotein transport activity determined by LF in ( D ) male and ( E ) female capillaries. Graphs of LF measuring P-glycoprotein (P-gp) transport activity in male ( F ) and female ( G ) capillaries (capillary N = 15–20 per experimental data point) exposed to 0.001 μM erastin over time. Dotted red line denotes multiple comparison. Significance is as compared to control unless otherwise specified: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: PSC833 (valspodar), the P-glycoprotein-specific inhibitor, was purchased from Tocris Bioscience.

Techniques: Activity Assay, Incubation, Fluorescence, Control, Comparison