kca3 1 channels tram 34  (Tocris)

Journal: Bioactive Materials

Article Title: Ion channel/Stat6-driven nano-immune programming of tissue-resident macrophages by amide-functionalized nanocellulose

doi: 10.1016/j.bioactmat.2026.03.038

Figure Lengend Snippet: Bulk RNA-Seq analysis of RAW 264.7 cells in vitro . (a) Unsupervised hierarchical clustering of DEGs (Log2FC, ∗ p < 0.05) associated with the immune response in RAW 264.7 cells after 14 days across various groups. (b – d) Representative KEGG enrichment analysis of key DEGs associated with biological process (BP), cellular component (CC), and molecular function (MF) in RAW 264.7 cells. (e, f) qRT-PCR validation of canonical M1 and M2 macrophage-specific gene markers expression in various groups at day 14 ( n = 5 each). (g) Unsupervised hierarchical clustering of DEGs (Log2FC, ∗ p < 0.05) associated with ion channel activity in RAW 264.7 cells after 14 days in various groups. (h) Pearson's correlation analysis of key DEGs associated with ion channel activity across groups. (i) Principal component analysis (PCA) analysis of variance in various treatment groups at day 14. (j, k) Fold change expression of Scn1b and KCa3.1 (= Kcnn4 ) (Log2FC, ∗ p < 0.05) associated with ion channel activity ( n = 5 each). (l, m) Representative immunostaining results for KCa3.1 (red) and Scn1b (blue) in various groups at day 14. Scale bar: 10 μm ( n = 5 each). (n, o) Immunostaining results with spatial fluoresence intensities for KCa3.1 and Stat6 in RAW 264.7 after ion channel inhibition using nifedipine (5 μM, L-type Ca 2+ ion channel blocker), TRAM-34 (10 μM, Ca 2+ -activated K + blocker), and AS1810722 (10 μM, Stat6 blocker) at day 14. Scale bar: 10 μm. Data reported as mean ± s.d. of replicated experiments ( n = 5 each), statistical significance was considered at ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 (One-way ANOVA test with Tukey's test post hoc analysis).

Article Snippet: For this, the RAW 264.7 cells (∼1.5 × 10 5 /mL/6-well) cultured with a-CNCs (100 μg mL −1 ) and ion channel inhibitors, such as Nifedipine (50 μM, Sigma-Aldrich), TRAM-34 (100 μM, MedChemExpress), and AS1810722 (100 μM, MedChemExpress) for 14 days.

Techniques: RNA Sequencing, In Vitro, Quantitative RT-PCR, Biomarker Discovery, Expressing, Activity Assay, Immunostaining, Inhibition

Journal: Redox Biology

Article Title: Physiological oxygen levels reset K + channel activity in human vascular endothelial cells

doi: 10.1016/j.redox.2025.103981

Figure Lengend Snippet: K + channel contributions to whole-cell current vary between cells adapted to 5 kPa or 18 kPa O 2 . ( A) Raw current traces under indicated conditions recorded from HUVEC exhibit outward rectifying whole-cell currents. ( B) Comparisons of normalised current amplitudes (relative to current at +50 mV) under indicated conditions show current inhibition relative to basal control from cells cultured at 5 kPa or 18 kPa using 10 mM TEA, 5 μM TRAM-34, 100 nM apamin (5 kPa: Basal vs. TEA: P = 0.0381, Basal vs. TEA + TRAM-34: P = 0.0095, Basal vs. TEA + TRAM-34+apamin: P = 0.0022; 18 kPa: Basal vs. TEA: P = 0.03723, Basal vs. TEA + TRAM-34: P = 0.0076, Basal vs. TEA + TRAM-34+apamin: P = 0.0042; two-way ANOVA, data denote mean ± S.E.M.). ( C ) Steady-state current-voltage (IV) relationships showing differential effects of TEA, TRAM-34 and apamin in HUVEC adapted to either O 2 level (5 kPa O 2 : F Treatment (3, 90) = 49.88, P < 0.0001, F Voltage (12, 90) = 131.0, P < 0.0001; 18 kPa O 2 : F Treatment (3, 104) = 15.50, P < 0.0001, F Voltage (12, 104) = 55.47, P < 0.0001, n = 2–3 cells each, two-way ANOVA, data denote mean (+S.E.M.).

Article Snippet: TRAM-34 (5 μM) was from Cambridge Bioscience Ltd, UK.

Techniques: Inhibition, Control, Cell Culture

Journal: Redox Biology

Article Title: Physiological oxygen levels reset K + channel activity in human vascular endothelial cells

doi: 10.1016/j.redox.2025.103981

Figure Lengend Snippet: K + channel contributions to whole-cell current vary between cells adapted to 5 kPa or 18 kPa O 2 . ( A) Raw current traces under indicated conditions recorded from hCMEC/D3 cells exhibit outward rectifying whole-cell currents. ( B ) Comparisons of normalised current amplitudes (relative to current at +50 mV) under indicated conditions show current inhibition relative to basal control from cells cultured at 5 kPa or 18 kPa using 10 mM TEA, 5 μM TRAM-34, 100 nM apamin (5 kPa: Basal vs. TEA: P = 0.0375, Basal vs. TEA + TRAM-34: P = 0.0013, Basal vs. TEA + TRAM-34+apamin: P < 0.0001; 18 kPa: Basal vs. +TEA + TRAM-34+apamin: P = 0.0244, two-way ANOVA, data denote mean ± S.E.M.). ( C ) Steady-state current-voltage (IV) relationships showing differential effects of TEA, TRAM-34 and apamin in hCMEC/D3 cells adapted to either O 2 level (5 kPa O 2 : F Treatment (3, 102) = 173.0, P < 0.0001, F Voltage (12, 102) = 131.4, P < 0.0001; 18 kPa O 2 : F Treatment (3, 103) = 14.96, P < 0.0001, F Voltage (12, 103) = 38.59, P < 0.0001, n = 3 cells each, two-way ANOVA, data denote mean (+S.E.M.).

Article Snippet: TRAM-34 (5 μM) was from Cambridge Bioscience Ltd, UK.

Techniques: Inhibition, Control, Cell Culture