Journal: Scientific reports

Article Title: High-volume hemofiltration does not protect human kidney endothelial and tubular epithelial cells from septic plasma-induced injury.

doi: 10.1038/s41598-024-69202-z

Figure Lengend Snippet: Figure 7. Evaluation of TEC polarity after incubation with plasma collected at different HVHF/SVHF time points. (A, B) Evaluation of cellular polarity by analysis of trans-epithelial electrical resistance (TEER in A) and FITC-albumin reabsorption (B) of TEC incubated with plasma collected at different time points after HVHF (black columns) or SVHF (white columns) start. In respect to Vehicle alone, d1h0 plasma induced a significant decrease of TEER and albumin reabsorption (*p < 0.05 d1h0 vs. Vehicle). Plasma samples collected at d1h6 and d1h12 increased TEC polarity (°p < 0.05 d1h6 or d1h12 vs. d1h0). By contrast, TEC monolayers incubated with d3h0 plasma showed a new significant decrease of both TEER and albumin reabsorption (#p < 0.05 d3h0 vs. d1h6 or d1h12). Similar results were observed for HVHF and SVHF. All experiments were performed in triplicate. (C, D) Representative micrographs (C) and relative fluorescence intensity quantification (D) of Megalin (green staining), NKCC1 (green staining) and SGLT-2 (red staining) expression in TEC incubated with plasma collected at different time points after HVHF start. In respect to Vehicle alone, d1h0 plasma induced a significant decrease of Megalin, NKCC1 and SGLT-2 expression (*p < 0.05 d1h0 vs. Vehicle). Plasma samples collected at d1h6 and d1h12 increased the staining for all the proteins on TEC surface (°p < 0.05 d1h6 or d1h12 vs. d1h0). By contrast, TEC monolayers incubated with d3h0 plasma showed a new significant decrease of Megalin, NKCC1 and SGLT-2 expression (#p < 0.05 d3h0 vs. d1h6 or d1h12). Similar results were observed using SVHF plasma. All experiments were performed in triplicate. In fluorescence micrographs, nuclei were counterstained in blue by Hoechst.

Article Snippet: After appropriate stimuli, cultured TEC were fixed in ethanol/acetic acid 2:1 and stained with antibodies directed to human Fas (Upstate Biotechnology, Lake Placid, NY, USA), megalin, NKCC1 or SGLT-2 (all from Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Incubation, Clinical Proteomics, Fluorescence, Staining, Expressing

Journal: The Journal of Experimental Medicine

Article Title: Immunization with vaccinia virus induces polyfunctional and phenotypically distinctive CD8 + T cell responses

doi: 10.1084/jem.20062363

Figure Lengend Snippet: Vaccinia virus–specific CD8 + T cells express intracellular granzymes and perforin. Perforin and granzyme expression on KVD tetramer-binding cells is shown as red contour plots overlayed on density plots of the total CD8 + T cell population. An APC-conjugated tetramer was used in A; a PE-conjugated tetramer was used in B. Representative staining from one of three donors.

Article Snippet: Cells were permeablized using the Foxp3 Staining Buffer Set (eBioscience), according to manufacturer's instructions, and intracellularly stained with antibodies against granzyme A–PE or –FITC (BD Biosciences), perforin-FITC (BD Biosciences), and granzyme B–APC (Caltag).

Techniques: Expressing, Binding Assay, Staining

Journal: medRxiv

Article Title: Urinary Astrocyte-derived Extracellular Vesicles: A Non-invasive Tool for Capturing Human In Vivo Molecular “Movies” of Brain

doi: 10.1101/2024.01.12.24301104

Figure Lengend Snippet: (a) Schematic diagram of the uADEVs isolation protocol. (b and e) NTA results of uTEVs and uADEVs. (c, d, f, and g) TEM images of uTEVs and uADEVs (scale bars: 0.5 μm and 100nm). (h) Results of western blotting: Three EV markers (CD63, CD9 and Alix) and an astrocyte marker (GFAP) were present in the ADEVs sample, while two kidney markers (NKCC2 and NCC) were absent. (i) Significantly increased astrocyte-related neurotrophic factors in uADEVs.)

Article Snippet: NKCC2: Anti-NKCC2 antibody (Abcam, Cat No: ab171747) at 1/2000 dilution, protein loading amount: 4 μg per lane.

Techniques: Isolation, Western Blot, Marker