calcium channel blockers verapamil  (MedChemExpress)

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Disruption of cellular calcium homeostasis by duck Tembusu virus facilitates viral replication via AMPK pathway activation

doi: 10.3389/fcimb.2026.1743907

Figure Lengend Snippet: DTMUV infection increases cytoplasmic Ca 2+ levels in DEFs. (A) Flow cytometry profiles showing cytoplasmic Ca2+ levels in DEFs with and without DTMUV infection and probed with Flou-4AM. (B) Cytoplasmic Ca2+ levels of DEFs with (red) and without (blue) DTMUV infection (MOI = 0.1) for 6, 8, 10, or 12 hours, expressed as mean fluorescence intensity (MFI). (C) Cytoplasmic Ca2+ levels of DEFs with and without DTMUV infection and concurrent treatment with DMSO (control) verapamil or diltiazem hydrochloride. Data expressed as mean ± standard deviation (n = 3), analyzed using Student’s t-test; *p < 0.05, **p < 0.01, ****P<0.0001.

Article Snippet: The calcium channel blockers verapamil (Cat# HY-14275) and diltiazem hydrochloride (Cat# HY-14656), the AMPK inhibitor Compound C (Cat# HY-13418A), and calcium chelating agent BAPTA-AM (Cat# HY-100545) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Infection, Flow Cytometry, Fluorescence, Control, Standard Deviation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Disruption of cellular calcium homeostasis by duck Tembusu virus facilitates viral replication via AMPK pathway activation

doi: 10.3389/fcimb.2026.1743907

Figure Lengend Snippet: VDCC blockers and a cytoplasmic Ca 2+ chelator reduce DTMUV particle production. (A, B) Analysis of plaque assays of DEFs infected with DTMUV and treated with verapamil (25 µM), diltiazem hydrochloride (50 µM), or DMSO (control; (A) ), and BAPTA-AM (25 µM) or DMSO (control; (B) ). Results expressed as the viral titer ratio (%) between each drug-treated group and the control group at 12, 24, and 36 hpi. Data expressed as mean ± standard deviation of triplicate samples, analyzed by two-way ANOVA with multiple comparisons. *p < 0.05, **p <0.01, ***p <0.001, ****p < 0.0001. Results shown are representative of three independent experiments.

Article Snippet: The calcium channel blockers verapamil (Cat# HY-14275) and diltiazem hydrochloride (Cat# HY-14656), the AMPK inhibitor Compound C (Cat# HY-13418A), and calcium chelating agent BAPTA-AM (Cat# HY-100545) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Infection, Control, Standard Deviation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Disruption of cellular calcium homeostasis by duck Tembusu virus facilitates viral replication via AMPK pathway activation

doi: 10.3389/fcimb.2026.1743907

Figure Lengend Snippet: VDCC blockers and a cytoplasmic Ca 2+ chelator inhibit the replication step of DTMUV infection. (A) Viral entry assay of DEFs pretreated with DMSO, diltiazem (50 µM), or BAPTA-AM (25 µM) for 1 hour prior to DTMUV infection (MOI = 1) at 4°C for 1 hour and fusion at 37°C. Viral RNA levels in the cytoplasm were quantified by RT-qPCR at 2 hours post-infection (hpi), expressed as relative DTMUV mRNA levels between the drug-treated groups and the control group. (B) Viral replication assay of DEFs infected with DTMUV (MOI = 1) prior to treatment with DMSO (control), diltiazem hydrochloride (50 µM), or EAPTA-AM (25 µM) at 2 hpi, and RT-qPCR analysis of viral RNA replication in infected cells at 6 hpi, expressed as relative DTMUV mRNA levels between the drug-treated and control groups. (C) Plaque assay of viral release in DEFs cultured infected with DTMUV (MOI = 1) prior to treatment with DMSO (control), diltiazem hydrochloride (50 µM), or EAPTA-AM (25 µM) at 10 hpi and plating at 12 hpi. Results expressed as the viral titer ratio (%) between the drug-treated groups and the control group. (D) Viral replication assay of DEFs infected with DTMUV (MOI = 1) prior to treatment with DMSO (control) or alternative forms of verapamil (25 µM), diltiazem hydrochloride (50 µM), or BAPTA-AM (25 µM) at 1 hpi. Infected cells were harvested for RT-qPCR analysis of DTMUV mRNA levels at 8, 10, and 12 hpi, expressed as relative DTMUV mRNA levels between the drug-treated and control groups. Data expressed as mean ± standard deviation of triplicate samples, analyzed by one-way or two-way ANOVA with multiple comparisons; *p < 0.05, **p <0.01, ***p <0.001, ****p <0.0001. Data shown are representative of three independent experiments. ns: no significant difference.

Article Snippet: The calcium channel blockers verapamil (Cat# HY-14275) and diltiazem hydrochloride (Cat# HY-14656), the AMPK inhibitor Compound C (Cat# HY-13418A), and calcium chelating agent BAPTA-AM (Cat# HY-100545) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Infection, Quantitative RT-PCR, Control, Viral Replication Assay, Plaque Assay, Cell Culture, Standard Deviation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Disruption of cellular calcium homeostasis by duck Tembusu virus facilitates viral replication via AMPK pathway activation

doi: 10.3389/fcimb.2026.1743907

Figure Lengend Snippet: DTMUV-mediated AMPK activation is markedly diminished by treatment with VDCC blockers or a cytoplasmic Ca 2+ chelator. (A) Western blotting of pAMPKα (Thr172) in DEFs infected with DTMUV (MOI = 1) and harvested at the indicated time points. (B) Immunoblotting analysis of pAMPKα (Thr172) levels in DEFs treated with DMSO (control), verapamil (25 µM; Vera), diltiazem hydrochloride (50 µM; Dilt), or BAPTA-AM (25 µM; BAP), with and without DTMUV infection (MOI = 1) for 12 hours.

Article Snippet: The calcium channel blockers verapamil (Cat# HY-14275) and diltiazem hydrochloride (Cat# HY-14656), the AMPK inhibitor Compound C (Cat# HY-13418A), and calcium chelating agent BAPTA-AM (Cat# HY-100545) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Activation Assay, Western Blot, Infection, Control

Journal: Advanced Science

Article Title: Magnetically‐Induced Suppression of Oxidative Stress Prevents Venous Thrombosis

doi: 10.1002/advs.202513299

Figure Lengend Snippet: Pharmacological inhibition of the Ca 2+ and NOX4 enzyme largely diminished the protective effect of SMF, supporting the involvement of the Ca 2+ ‐ATP‐NOX4 axis. A) Intracellular ATP levels were quantified in C166 cells treated with or without Calcium channel blocker (Carboxyamidotriazole, CAI) and SMF for 24 h, followed by H 2 O 2 stimulation for 2 h (n = 53). B) NOX4 enzyme activity was detected by the H 2 O 2 level change in C166 cells pretreated with CAI for 24 h and SMF, then exposed to basal or H 2 O 2 ‐stimulated conditions. NOX4 enzyme activity was assessed by its production of H 2 O 2 using a fluorometric assay (The absolute‐quantitative: the absolute concentration of H 2 O 2 can be calculated from a standard curve, Elabscience H 2 O 2 Assay Kit, and semi‐quantitative methods with Amplex Red fluorescence assay). C,D) Representative immunofluorescence images of ROS (green) C) and Flow cytometry quantification of intracellular ROS levels D) in C166 cells subjected to 24‐h CAI pretreatment and SMF exposure following 0.4 m m H 2 O 2 exposure (IF for DCFH‐DA probe staining, five 200 × FOV were randomly recorded and analyzed, FCM for DCFH‐DA fluorescence intensity, n = 3). E) Cell viability of C166 cells subjected to 0.4 m m H 2 O 2 with or without 24‐h SMF exposure and CAI pretreatment (n = 5). F,G) Representative immunofluorescence images of ROS (F) and flow cytometry quantification of intracellular ROS levels(G) in C166 cells subjected to 0.4 m m H 2 O 2 following 24‐h NOX4 enzyme inhibition (GLX351322) and SMF exposure (five 200 × FOV were randomly recorded and analyzed, n = 3). H) Cell viability of C166 cells subjected to 0.4 m m H 2 O 2 with or without 24‐h GLX351322 pretreatment and SMF exposure (n = 5). Data represent the means ± SD from three or five independent experiments. Statistical significance was determined by one‐way ANOVA followed by Fisher's LSD test for the indicated pairs. ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Appropriate concentrations of the calcium channel blocker Carboxyamidotriazole (MedChemExpress, Cat# HY‐16126) were determined using CCK8 and the Fluo‐4 Calcium Assay Kit.

Techniques: Inhibition, Activity Assay, Concentration Assay, Fluorescence, Immunofluorescence, Flow Cytometry, Staining, Enzyme Inhibition Assay