l type calcium channel agonist  (Alomone Labs)

Journal: International Journal of Molecular Sciences

Article Title: AsHC 360 Exposure Influence on Epileptiform Discharges in Hippocampus of Infantile Male Rats In Vitro

doi: 10.3390/ijms242316806

Figure Lengend Snippet: EDs within the CA3 region of hippocampal slices under 10 nM (S)-(-)-Bay-K-8644 and 100 μg As L −1 AsHC 360 co-exposure. ( A ) Long-term records and representative ED clusters. The boxplots of IIDs and IDs ( B ) amplitude, ( C ) frequency, and ( D ) duration within Pre-, Adding, and Post-stages. “ns” represents no significant difference, “*” indicates p < 0.05, and “***” indicates p < 0.001.

Article Snippet: L-type calcium channel agonist (S)-(-)-Bay-K-8644 was obtained from MCE (Med Chem Express LLC, Shanghai, China).

Techniques:

Journal: PLoS ONE

Article Title: Suppression of Protective Responses upon Activation of L-Type Voltage Gated Calcium Channel in Macrophages during Mycobacterium bovis BCG Infection

doi: 10.1371/journal.pone.0163845

Figure Lengend Snippet: PMA stimulated THP1 macrophages were either infected with 2 MOI M . bo vis BCG (BCG) or stimulated with 50 nM BAYK8644 or both for indicated times. 30 min before the incubation period, cells were incubated with 10 μM DCFH-DA. After the incubation period, cells were washed with culture medium and ROS levels were analyzed by flow cytometry. In all the Panels, thin line represents ROS generation in uninfected or unstimulated or control cells; thick line represents ROS generation in infected or stimulated cells as indicated. Bar graphs adjacent to histograms in Panel (A-C) show Mean Fluorescent Intensity (MFI) of the peak at the higher fluorescence in the figure. Data from one of three independent experiments are shown (n = 3). The star above the bars represents the P value between control and the corresponding group of that bar in each panel. The results were analyzed by one way ANOVA followed by Tukey’s post hoc multiple comparison test. * = P ≤ 0.05; ** = P ≤ 0.01; *** = P ≤ 0.001 and **** = P ≤ 0.0001.

Article Snippet: L-type calcium channel agonist BAYK8644 [ ] and antagonist Amlodipine [ ] were purchased from Tocris (United Kingdom).

Techniques: Infection, Incubation, Flow Cytometry, Control, Fluorescence, Comparison

Journal: PLoS ONE

Article Title: Suppression of Protective Responses upon Activation of L-Type Voltage Gated Calcium Channel in Macrophages during Mycobacterium bovis BCG Infection

doi: 10.1371/journal.pone.0163845

Figure Lengend Snippet: For Panel A, macrophages were derived from bone marrow of female Balb/c mice. For Panel B, PBMCs from healthy volunteers were differentiated into macrophages. Macrophages were either infected with 2 MOI M . bovis BCG or stimulated with 50 nM BAYK8644 or both for 1h and ROS was estimated as described in . In both the Panels, thin line represents ROS generation in uninfected or unstimulated or control cells; thick line represents ROS generation in infected or stimulated cells as indicated. Bar chart below each panel represents MFI of the peak at the higher fluorescence in the figure. Data from one of three independent experiments are shown (n = 3). The star above the bar represents the P value between control and the corresponding group of that bar in each panel. The results were analyzed by one way ANOVA followed by Tukey’s post hoc multiple comparison test. * = P ≤ 0.05; ** = P ≤ 0.01; *** = P ≤ 0.001 and **** = P ≤ 0.0001.

Article Snippet: L-type calcium channel agonist BAYK8644 [ ] and antagonist Amlodipine [ ] were purchased from Tocris (United Kingdom).

Techniques: Derivative Assay, Infection, Control, Fluorescence, Comparison

Journal: PLoS ONE

Article Title: Suppression of Protective Responses upon Activation of L-Type Voltage Gated Calcium Channel in Macrophages during Mycobacterium bovis BCG Infection

doi: 10.1371/journal.pone.0163845

Figure Lengend Snippet: PMA simulated THP1 cells were transfected with siRNAs against indicated molecules for 36 h followed by infection with 2 MOI M . bovis BCG (Panel A) and/or stimulation with 50 nM BAYK8644 (Panel B) or both (Panel C) for 1 h and ROS was estimated as described above. In all the panels, thin line represents ROS generation in unstimulated/uninfected or control cells transfected with control siRNAs. Dotted line represent ROS generation in cells transfected with control siRNA followed by either infection with M . bovis BCG (Panel A) or stimulation with BAYK8644 (Panel B) or both (Panel C). The thick line in all the panels represents ROS generation in cells either infected with M . bovis BCG (Panel A) or stimulation with BAYK8644 (Panel B) or both (Panel C) following transfection with specific siRNAs against indicated molecules. Bar graph below each panel represents MFI of the peak at the higher fluorescence in the figure. Data from one of three independent experiments are shown (n = 3). The star above the bar represents the P value between stimulated/infected groups transfected with control siRNAs and the corresponding group of that bar in each panel. The results were analyzed by one way ANOVA followed by Tukey’s post hoc multiple comparison test. ns = P > 0.05; * = P ≤ 0.05; ** = P ≤ 0.01; *** = P ≤ 0.001 and **** = P ≤ 0.0001.

Article Snippet: L-type calcium channel agonist BAYK8644 [ ] and antagonist Amlodipine [ ] were purchased from Tocris (United Kingdom).

Techniques: Transfection, Infection, Control, Fluorescence, Comparison

Journal: PLoS ONE

Article Title: Suppression of Protective Responses upon Activation of L-Type Voltage Gated Calcium Channel in Macrophages during Mycobacterium bovis BCG Infection

doi: 10.1371/journal.pone.0163845

Figure Lengend Snippet: PMA stimulated THP1 macrophages were treated with inhibitors to indicated molecules for 1 h followed by either infection with 2 MOI M . bovis BCG or stimulation with 50 nM BAYK8644 or both for 1 h. In all the panels, thin line represents ROS generation in unstimulated/uninfected or control cells. Dotted line represent ROS generation in cells infected with M . bovis BCG (Panel A) or stimulation with BAYK8644 (Panel B) or both (Panel C). The thick line in all the panels represent ROS generation in cells either infected with M . bovis BCG (Panel A) or stimulated with BAYK8644 (Panel B) or both (Panel C) following treatment with inhibitors to indicated molecules. Bar graph below each panel represents MFI of the peak at the higher fluorescence in the figure. Data from one of three independent experiments are shown (n = 3). The star above the bar represents the P value between stimulated/infected cells and the corresponding group of that bar chart in each panel. The results were analyzed by one way ANOVA followed by Tukey’s post hoc multiple comparison test. ns = P > 0.05; * = P ≤ 0.05; ** = P ≤ 0.01; *** = P ≤ 0.001 and **** = P ≤ 0.0001.

Article Snippet: L-type calcium channel agonist BAYK8644 [ ] and antagonist Amlodipine [ ] were purchased from Tocris (United Kingdom).

Techniques: Infection, Control, Fluorescence, Comparison

Journal: PLoS ONE

Article Title: Suppression of Protective Responses upon Activation of L-Type Voltage Gated Calcium Channel in Macrophages during Mycobacterium bovis BCG Infection

doi: 10.1371/journal.pone.0163845

Figure Lengend Snippet: PMA stimulated THP-1 cells were transfected with siRNAs against indicated molecules for 36 h followed by infection with 2 MOI M . bovis BCG and/or stimulation with 50nM for 1 h and ROS was estimated as described above. In all the panels thin line represents ROS generation in unstimulaed/uninfected cells transfected with control siRNAs. Dotted line represent ROS generation in cells transfected with control siRNA followed by either infection with M . bovis BCG (Panel A) or stimulation with BAYK8644 (Panel B) or both (Panel C). The thick line in all the panels represent ROS generation in cells either infected with M . bovis BCG (Panel A) or stimulated with BAYK8644 (Panel B) or both (Panel C) following transfection with specific siRNAs against indicated molecules. Bar graph below each panel represents MFI of the peak at higher fluorescence in the figure. Data from one of the three independent experiments are shown (n = 3). The star above the bar represents the P value between unstimulated and the corresponding group of that bar in each panel. The results were analyzed by one way ANOVA followed by Tukey’s post hoc multiple comparison test. * = P < 0.05; ** = P < 0.01; *** = P < 0.001 and **** = P < 0.0001.

Article Snippet: L-type calcium channel agonist BAYK8644 [ ] and antagonist Amlodipine [ ] were purchased from Tocris (United Kingdom).

Techniques: Transfection, Infection, Control, Fluorescence, Comparison

Journal: PLoS ONE

Article Title: Suppression of Protective Responses upon Activation of L-Type Voltage Gated Calcium Channel in Macrophages during Mycobacterium bovis BCG Infection

doi: 10.1371/journal.pone.0163845

Figure Lengend Snippet: Either PMA stimulated THP1 macrophages or mouse bone marrow derived macrophages or human PBMC derived macrophages were either infected with 2 MOI M . bovis BCG or stimulated with 50 nM BAYK8644 or both for 24 h. Cytoplasmic extracts were western blotted for indicated molecules in the apoptotic (Panel A) or autophagic (Panel B) pathways. ‘Control’ represents unstimulated/uninfected cells. Numbers below the blots indicate relative intensities of the bands normalized with the housekeeping molecule GAPDH. All panels show one of three independent experiments (n = 3).

Article Snippet: L-type calcium channel agonist BAYK8644 [ ] and antagonist Amlodipine [ ] were purchased from Tocris (United Kingdom).

Techniques: Derivative Assay, Infection, Western Blot, Control

Journal: PLoS ONE

Article Title: Suppression of Protective Responses upon Activation of L-Type Voltage Gated Calcium Channel in Macrophages during Mycobacterium bovis BCG Infection

doi: 10.1371/journal.pone.0163845

Figure Lengend Snippet: PMA treated THP1 macrophages (Panel A) and mouse bone marrow derived macrophages (Panel B) were seeded on the coverslip and washed with RPMI 1640 medium and incubated in OPTIMEM medium with or without BAYK8644 for 1 h followed by infection with FM4-64 labeled M . bovis BCG (panel A) or GFP expressing M . tb H37Rv (Panel B) for 4 h. thirty minutes prior to the end of infection period, cells were incubated with 50nM of Lysotracker Green (for Panel A) or Lysotracker Green (for Panel B). At the end of incubation period cells were washed once with PBS and fixed with 4% paraformaldehyde for 1h. Following through washes, the cover slips were mounted with anti-fade containing DAPI. Confocal microscopy was performed on Leica TCS SP-8 confocal instrument, LAX Version 1.8.1.137. Bar chart in both Panel A and Panel B represents percentage of co-localization as determined by LAS AF Version 2.6.0 build 7266 of Leica Micro Systems CMS GmbH. Bars represent percentage of co-localization of the indicated groups of three independent experiments (n = 3). The stars represent the P value between unstimulated and corresponding stimulated (Bay/Amlodipine) group of that bar in each panel. The results were analyzed by one way ANOVA followed by Tukey’s post hoc multiple comparison test. * = P < 0.05; ** = P < 0.01; *** = P < 0.001 and **** = P < 0.0001.

Article Snippet: L-type calcium channel agonist BAYK8644 [ ] and antagonist Amlodipine [ ] were purchased from Tocris (United Kingdom).

Techniques: Derivative Assay, Incubation, Infection, Labeling, Expressing, Confocal Microscopy, Comparison

Journal: PLoS ONE

Article Title: Suppression of Protective Responses upon Activation of L-Type Voltage Gated Calcium Channel in Macrophages during Mycobacterium bovis BCG Infection

doi: 10.1371/journal.pone.0163845

Figure Lengend Snippet: Bone marrow derived mouse macrophages (Panel A&B) either were infected with 2 MOI of M . bovis BCG or stimulated with 50nM of BAYK8644 or both for 24 h. For Panel A, culture supernatant was processed for the estimation of indicated cytokines and bars represent the amount of cytokine in pg/ml. For Panel B, cells were processed for measuring surface densities of indicated cytokine receptors by FACS and bars represent MFI of indicated groups of three independent experiments (n = 3). The results were analyzed by one way ANOVA followed by Tukey’s post hoc multiple comparison test. The star above the bar represents the P value between unstimulated/uninfected or control and the corresponding group of that bar in each panel. ns = P>0.05; * = P < 0.05; ** = P < 0.01; *** = P < 0.001 and **** = P < 0.0001.

Article Snippet: L-type calcium channel agonist BAYK8644 [ ] and antagonist Amlodipine [ ] were purchased from Tocris (United Kingdom).

Techniques: Derivative Assay, Infection, Comparison, Control

Journal: PLoS ONE

Article Title: Suppression of Protective Responses upon Activation of L-Type Voltage Gated Calcium Channel in Macrophages during Mycobacterium bovis BCG Infection

doi: 10.1371/journal.pone.0163845

Figure Lengend Snippet: Human PBMC derived macrophages (Panel A&B) either were infected with 2 MOI M . bovis BCG or stimulated with 50 nM BAYK8644 or both for 24 h. For Panel A, culture supernatant was processed for the estimation of indicated cytokines and bars represent the amount of cytokine in pg/ml. For Panel B, cells were processed for measuring surface densities of indicated cytokine receptors by FACS and bars represent MFI of indicated groups of three independent experiments (n = 3). The results were analyzed by one way ANOVA followed by Tukey’s post hoc multiple comparison test. The star above the bar represents the P value between unstimulated/uninfected and the corresponding group of that bar in each panel. ns = P>0.05; * = P < 0.05; ** = P < 0.01; *** = P < 0.001 and **** = P < 0.0001.

Article Snippet: L-type calcium channel agonist BAYK8644 [ ] and antagonist Amlodipine [ ] were purchased from Tocris (United Kingdom).

Techniques: Derivative Assay, Infection, Comparison