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: Biomaterials

Article Title: A bioprinted and scalable model of human tubulo-interstitial kidney fibrosis.

doi: 10.1016/j.biomaterials.2024.123009

Figure Lengend Snippet: Fig. 1. Generation of stable human kidney cell lines with proven origin. A: Schematic of the human kidney and the cortex illustrates the source of isolated cells. B: Immunofluorescence staining of PDGFRβ+, CD10+ and CD31+ specific antibodies for cells in cortex of human kidney. C: Enumeration of the workflow from isolating the cells to the generation of genetically tagged cells. D: Representative pictures of genetically labeled immortalized PDGFRβ+, proximal tubular epithelial and endothelial cell lines. 4′,6-diamidino-2-phenylindole (DAPI) was used for nuclear staining. E: Principal component analysis (PCA) plot of the generated three cell lines. The three cell lines show higher variability from each other, while technical replicates are similar to each other. F: Shared DE genes between isolated cell lines and human bulk RNA seq [13]. G: CD10+ (red) cells stained for cell-specific proximal tubule epithelial markers cubilin (green, left) and CD13 (green, right) including DAPI. H: Heatmap of bulk RNA-sequencing data demonstrating the genes expressed in CD10+. I: P-gp Calcein-AM transporter assay of CD10+ and negative control PDGFRβ+ cells, with or without transporter inhibitor PSC833. ***p < 0.001 (one-way ANOVA analysis followed by Tukey post-test), n = 3 with each 12 replicates. J: Presence of intracellular vWF (red) in CD31+ (green) cells including DAPI. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Cells were treated for 1 h with 5 μM P-gp transporter inhibitor PSC833 (Tocris Biosciences, Bristol, UK) in KH buffer at 37 ◦C, 5 % (v/v) CO2, and afterwards exposed for 1 h to 1 μM Calcein-AM (Life Technologies Europe BV) with or without 5 μM PSC833 in KH buffer.

Techniques: Isolation, Immunofluorescence, Staining, Labeling, Generated, RNA Sequencing, Negative Control