Journal: Cell Death Discovery

Article Title: Therapeutic potential of voltage-dependent potassium channel subtype 1.3 blockade in alleviating macrophage-related renal inflammation and fibrogenesis

doi: 10.1038/s41420-025-02508-7

Figure Lengend Snippet: A Respective western blotting image and densitometry analysis of ERK/NF-κB signaling after transfection with LV-Kv1.3 in THP-1 cells. Data were expressed as mean ± SD (n = 4). B The phosphorylation signal data are depicted in the form of stacked histogram overlaid plots. The mean fluorescence intensity (MFI) of the ERK1/2 and NF-κB p65 phosphorylation signal was expressed as the mean ± SD (n = 3). C Effects of MgTx, PD98059, and Licochalcone B on Kv1.3 and ERK/NF-κB signaling in LV-Kv1.3-transduced THP-1 cells were analyzed by western blotting. Data were expressed as mean ± SD (n = 3). D Respective western blotting image and densitometry analysis of ERK/NF-κB signaling in HK-2 cells co-cultured with the supernatant from LV-Kv1.3-transduced THP-1 cells for 48 h. Data were expressed as mean ± SD (n = 4). E Respective western blotting image and densitometry analysis of ERK/NF-κB signaling in mice subjected to UUO (7 days post-surgery) and IRI. Data were expressed as mean ± SD (n = 4).

Article Snippet: MgTx, PD98059 and Licochalcone B were obtained from MedChemExpress (HY-P1280, HY-12028 and HY-N0373, respectively).

Techniques: Western Blot, Transfection, Phospho-proteomics, Fluorescence, Cell Culture

Journal: Cells

Article Title: Reduction of Ca 2+ Entry by a Specific Block of KCa3.1 Channels Optimizes Cytotoxic Activity of NK Cells against T-ALL Jurkat Cells.

doi: 10.3390/cells12162065

Figure Lengend Snippet: Figure 2. KCa3.1 and Kv1.3 currents in NK cells of healthy donors. (A–C) Examples of whole- cell current recordings in response to voltage ramps from −150 to +50 mV. A. Activated NK cell, mainly expressing NS6180-sensitive KCa3.1 current (KCa type). (B). Resting NK cell expressing only margatoxin-sensitive Kv1.3 current (Kv type). (C). Activated NK cells, expressing both KCa.1 and Kv1.3 currents (KCa + Kv). Dashed lines mark zero current level. (D). KCa3.1 single channel recording in the whole-cell mode. Steady state record at −100 mV (top). Unitary current–voltage relation (bottom), leak current was subtracted from recordings, displaying 1 or 2 open KCa3.1 channels. Slope conductance was calculated from linear regression (dashed line).

Article Snippet: Product Supplier Catalog/ Specification Number Stock Concentration Solvent Anti-CD107a-FITC Biolegend 326810 NS NS Fluo-4 AM Invitrogen by Thermo Fisher Scientific F1221 2 mM DMSO Fura-2 AM Invitrogen by Thermo Fisher Scientific F14201 2 mM DMSO CellTracker Deep Red Invitrogen by Thermo Fisher Scientific C34565 2 mM DMSO Hoechst 33342 Thermo FisherScientific 62249 20 mM H2O Blockers, salts, buffers Monensin Sigma-Aldrich 22373-78-0 2 mM Methanol NS6180 Tocris 4864 10 mM DMSO MgTx Tocris 3563 10 mM H2O PBS Gibco 70013-032 10X H2O CaCl2 Sigma C-7902 1 M H2O MgCl2 Sigma M-2670-100G 1 M H2O KCl Sigma P-5405 NA H2O EGTA Sigma E-4378 100 mM H2O HEPES Sigma H-7523 NA H2O KOH Sigma P-5958 1 M H2O NaOH Sigma S-0899 1 M H2O

Techniques: Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Induction of IL-17 and nonclassical T-cell activation by HIV-Tat protein

doi: 10.1073/pnas.1308673110

Figure Lengend Snippet: T-cell activation by Tat is dependent upon VEGFR2 and downstream signals. (A) SU 1498, a VEGFR2 inhibitor, inhibited Tat mediated T-cell stimulation. (B) T cells were transfected without RNA (mock) nonspecific siRNA (si-control) or siRNA to VEGFR2 (si-VEGFR2) for 24 h and then exposed to Tat for 24 h. (C) Pretreatment of T cells with TPCK (NFκB inhibitor), genistein (Gen; TK inhibitor), LY294002 (PI3 kinase inhibitor), SNS-032 (CDK 2, 7, and 9 inhibitor) or with rapamycin (Rapa; mTOR inhibitor) inhibited Tat-mediated T-cell stimulation. (D) Pretreatment of T cells with RO-32-0432 (PKC inhibitor) or margatoxin (MgTX; Kv1.3 inhibitor) had no effect on Tat-mediated T-cell stimulation. Data are mean ± SD from three to five independent experiments compared by ANOVA with Bonferroni’s multiple comparison, *P < 0.05, **P < 0.001; ns, not significant.

Article Snippet: The following pharmacological inhibitors were used for mapping of pathways: Tosyl phenylalanyl chloromethyl ketone (Sigma, 10 μM), RO-32–0432 (Sigma, 100 nM), Margatoxin (Santa Cruz, 100 nM), Genistein (Sigma, 100 μM), {"type":"entrez-nucleotide","attrs":{"text":"LY294002","term_id":"1257998346","term_text":"LY294002"}} LY294002 (Cell Signaling, 100 μM), SNS-032 (Selleck, 0.5 μM), Rapamycin (Selleck, 5 nM), Nocodazole (Sigma, 5 mg/mL), SU 1498 (Invitrogen, 2 μM), Chlorpromazine (Sigma, 30 μM), Amiloride (Sigma catalog, 500 μM), and Filipin (Sigma, 3 μg/mL).

Techniques: Activation Assay, Cell Stimulation, Transfection, Control, Comparison