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  • 94
    Alomone Labs snx 482
    Snx 482, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Synaptic Systems cav2 3
    Representative images and quantification of western blot analysis of calcium- and voltage-gated potassium channels. (A) Total expression of BK and SK2 potassium channels in mouse hippocampus showed no significant differences between WT and <t>Cav2.3-KO</t> mice (WT = 8, KO = 6; p > 0.05). In contrast, in the Cav2.3-KO, the total level of Kv4.2 was significantly increased compared to the WT (WT = 8, KO = 6; p
    Cav2 3, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Santa Cruz Biotechnology cacna1s
    Proposed model for <t>CACNA1S</t> expression by M . tb . Stimulation of macrophages by M . tb or Rv2463 increases CREB phosphorylation in a TLR dependent MyD88 independent pathway that involves IRAK1 and TRAF6. Phosphorylated CREB (p-CREB) translocates to the nucleus and binds to CACNA1S promoter to induce CACNA1S protein expression. In parallel mycobacterial stimulation attenuates the generation of reactive oxygen species (ROS) that further contributes to CREB phosphorylation. Calcium homeostasis sensors STIM1 and STIM2 also play an inhibitory role in CACNA1S upregulation upon M . tb infection or Rv2463 stimulation.
    Cacna1s, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Alomone Labs calciseptine
    Proposed model for <t>CACNA1S</t> expression by M . tb . Stimulation of macrophages by M . tb or Rv2463 increases CREB phosphorylation in a TLR dependent MyD88 independent pathway that involves IRAK1 and TRAF6. Phosphorylated CREB (p-CREB) translocates to the nucleus and binds to CACNA1S promoter to induce CACNA1S protein expression. In parallel mycobacterial stimulation attenuates the generation of reactive oxygen species (ROS) that further contributes to CREB phosphorylation. Calcium homeostasis sensors STIM1 and STIM2 also play an inhibitory role in CACNA1S upregulation upon M . tb infection or Rv2463 stimulation.
    Calciseptine, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Representative images and quantification of western blot analysis of calcium- and voltage-gated potassium channels. (A) Total expression of BK and SK2 potassium channels in mouse hippocampus showed no significant differences between WT and Cav2.3-KO mice (WT = 8, KO = 6; p > 0.05). In contrast, in the Cav2.3-KO, the total level of Kv4.2 was significantly increased compared to the WT (WT = 8, KO = 6; p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Functional Coupling of Cav2.3 and BK Potassium Channels Regulates Action Potential Repolarization and Short-Term Plasticity in the Mouse Hippocampus

    doi: 10.3389/fncel.2019.00027

    Figure Lengend Snippet: Representative images and quantification of western blot analysis of calcium- and voltage-gated potassium channels. (A) Total expression of BK and SK2 potassium channels in mouse hippocampus showed no significant differences between WT and Cav2.3-KO mice (WT = 8, KO = 6; p > 0.05). In contrast, in the Cav2.3-KO, the total level of Kv4.2 was significantly increased compared to the WT (WT = 8, KO = 6; p

    Article Snippet: The result shown in shows that in mouse hippocampus, Cav2.3 and BK channels form a macromolecular complex.

    Techniques: Western Blot, Expressing, Mouse Assay

    AP waveform properties from CA1 pyramidal neurons. (A) Average first AP elicited by a +150 pA current injection for WT (black traces) and Cav2.3 KO (red traces) ± SEM. (B–G) Quantification of (A) . Firing threshold = first derivative of voltage > 0.2, and AP onset = time between current step onset and firing threshold. After-hyperpolarization (AHP) is calculated relative to firing threshold. (H) Example traces showing the first six APs elicited by a +200 pA current injection for WT (black traces) and Cav2.3 KO (red traces). (I–N) Quantification of (H) . Instead of AP onset, this analysis shows the inter-spike interval (time between peak voltages) and AHP was calculated as the minimum voltage between two APs. n = 17 for WT, and 19 for KO, all data shown as mean ± SEM, with Student’s T -Test to probe for significance and Holm-Sidak to account for multiple comparisons (n.s. = not significant, * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Functional Coupling of Cav2.3 and BK Potassium Channels Regulates Action Potential Repolarization and Short-Term Plasticity in the Mouse Hippocampus

    doi: 10.3389/fncel.2019.00027

    Figure Lengend Snippet: AP waveform properties from CA1 pyramidal neurons. (A) Average first AP elicited by a +150 pA current injection for WT (black traces) and Cav2.3 KO (red traces) ± SEM. (B–G) Quantification of (A) . Firing threshold = first derivative of voltage > 0.2, and AP onset = time between current step onset and firing threshold. After-hyperpolarization (AHP) is calculated relative to firing threshold. (H) Example traces showing the first six APs elicited by a +200 pA current injection for WT (black traces) and Cav2.3 KO (red traces). (I–N) Quantification of (H) . Instead of AP onset, this analysis shows the inter-spike interval (time between peak voltages) and AHP was calculated as the minimum voltage between two APs. n = 17 for WT, and 19 for KO, all data shown as mean ± SEM, with Student’s T -Test to probe for significance and Holm-Sidak to account for multiple comparisons (n.s. = not significant, * p

    Article Snippet: The result shown in shows that in mouse hippocampus, Cav2.3 and BK channels form a macromolecular complex.

    Techniques: Injection

    Altered synaptic connectivity between CA1 and subiculum. (A–D) Analysis of spontaneous excitatory postsynaptic currents (sEPSCs) recorded from subicular pyramidal neurons in the presence of Gabazine. (A) Representative traces from WT (black traces) and Cav2.3 KO (red traces) and cumulative probability of sEPSC amplitudes for all recorded events ( n > 5,000). Insets represent the average traces over all sEPSCs. (B) Quantification of amplitude and frequency for the average event recorded from each cell per genotype ( n = 20). (C) Cumulative distribution and quantification for the rise- and decay-times for the average sEPSCs. (D) Quantification for the charge conveyed with each sEPSC (area under the current trace). (E–H) Experimental setup (E) and analysis (F–H) of paired-pulse facilitation between CA1 and subiculum. (F,G) EPSP amplitude and decay time are not significantly different between WT and KO [analyzed from the first elicited EPSP of the 1,000 ms inter-stimulus intervals (ISIs) pulse]. Insets in (H) represent average traces for WT (black) and Cav2.3 KO (red) for two representative ISI. All data shown as mean ± SEM, with student’s T -Test or two-way ANOVA where appropriate, to probe for significance and Holm-Sidak to account for multiple comparisons (n.s. = not significant, * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Functional Coupling of Cav2.3 and BK Potassium Channels Regulates Action Potential Repolarization and Short-Term Plasticity in the Mouse Hippocampus

    doi: 10.3389/fncel.2019.00027

    Figure Lengend Snippet: Altered synaptic connectivity between CA1 and subiculum. (A–D) Analysis of spontaneous excitatory postsynaptic currents (sEPSCs) recorded from subicular pyramidal neurons in the presence of Gabazine. (A) Representative traces from WT (black traces) and Cav2.3 KO (red traces) and cumulative probability of sEPSC amplitudes for all recorded events ( n > 5,000). Insets represent the average traces over all sEPSCs. (B) Quantification of amplitude and frequency for the average event recorded from each cell per genotype ( n = 20). (C) Cumulative distribution and quantification for the rise- and decay-times for the average sEPSCs. (D) Quantification for the charge conveyed with each sEPSC (area under the current trace). (E–H) Experimental setup (E) and analysis (F–H) of paired-pulse facilitation between CA1 and subiculum. (F,G) EPSP amplitude and decay time are not significantly different between WT and KO [analyzed from the first elicited EPSP of the 1,000 ms inter-stimulus intervals (ISIs) pulse]. Insets in (H) represent average traces for WT (black) and Cav2.3 KO (red) for two representative ISI. All data shown as mean ± SEM, with student’s T -Test or two-way ANOVA where appropriate, to probe for significance and Holm-Sidak to account for multiple comparisons (n.s. = not significant, * p

    Article Snippet: The result shown in shows that in mouse hippocampus, Cav2.3 and BK channels form a macromolecular complex.

    Techniques:

    Cav2.3 knockout (KO) CA1 pyramidal cells are hyperexcitable. (A) Resting membrane potential (Vrest; A1 ) and cell capacitance (A2) , measured after break-in. (B) Input resistance quantified as the slope of an IV-curve for hyperpolarizing current steps from −160 to −20 pA. Insets represent averaged voltage traces for wildtype (WT; black) and KO (red) recordings during current injection. (C) Hyperpolarization activated current I H , measured as sag-percentage from maximum voltage deflection (C1) and as the slope of the rebound potential after the end of a series of hyperpolarizing current injections plotted against the steady-state voltage during the current injection (C2) . (D) Firing frequency of action potentials (APs) in CA1 pyramidal neurons of Cav2.3 KO and WT animals, elicited by current steps from Vrest, by injection of positive or negative current in steps of 20 pA. Inset shows example recordings for injections of +200 pA WT (black traces) or Cav2.3 KO (red traces). n = 17 for WT, and 21 for KO. All data shown as mean ± SEM, with Student’s T -Test to probe for significance (n.s. = not significant, * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Functional Coupling of Cav2.3 and BK Potassium Channels Regulates Action Potential Repolarization and Short-Term Plasticity in the Mouse Hippocampus

    doi: 10.3389/fncel.2019.00027

    Figure Lengend Snippet: Cav2.3 knockout (KO) CA1 pyramidal cells are hyperexcitable. (A) Resting membrane potential (Vrest; A1 ) and cell capacitance (A2) , measured after break-in. (B) Input resistance quantified as the slope of an IV-curve for hyperpolarizing current steps from −160 to −20 pA. Insets represent averaged voltage traces for wildtype (WT; black) and KO (red) recordings during current injection. (C) Hyperpolarization activated current I H , measured as sag-percentage from maximum voltage deflection (C1) and as the slope of the rebound potential after the end of a series of hyperpolarizing current injections plotted against the steady-state voltage during the current injection (C2) . (D) Firing frequency of action potentials (APs) in CA1 pyramidal neurons of Cav2.3 KO and WT animals, elicited by current steps from Vrest, by injection of positive or negative current in steps of 20 pA. Inset shows example recordings for injections of +200 pA WT (black traces) or Cav2.3 KO (red traces). n = 17 for WT, and 21 for KO. All data shown as mean ± SEM, with Student’s T -Test to probe for significance (n.s. = not significant, * p

    Article Snippet: The result shown in shows that in mouse hippocampus, Cav2.3 and BK channels form a macromolecular complex.

    Techniques: Knock-Out, Injection

    Proposed model for CACNA1S expression by M . tb . Stimulation of macrophages by M . tb or Rv2463 increases CREB phosphorylation in a TLR dependent MyD88 independent pathway that involves IRAK1 and TRAF6. Phosphorylated CREB (p-CREB) translocates to the nucleus and binds to CACNA1S promoter to induce CACNA1S protein expression. In parallel mycobacterial stimulation attenuates the generation of reactive oxygen species (ROS) that further contributes to CREB phosphorylation. Calcium homeostasis sensors STIM1 and STIM2 also play an inhibitory role in CACNA1S upregulation upon M . tb infection or Rv2463 stimulation.

    Journal: PLoS ONE

    Article Title: Regulation of L-type Voltage Gated Calcium Channel CACNA1S in Macrophages upon Mycobacterium tuberculosis Infection

    doi: 10.1371/journal.pone.0124263

    Figure Lengend Snippet: Proposed model for CACNA1S expression by M . tb . Stimulation of macrophages by M . tb or Rv2463 increases CREB phosphorylation in a TLR dependent MyD88 independent pathway that involves IRAK1 and TRAF6. Phosphorylated CREB (p-CREB) translocates to the nucleus and binds to CACNA1S promoter to induce CACNA1S protein expression. In parallel mycobacterial stimulation attenuates the generation of reactive oxygen species (ROS) that further contributes to CREB phosphorylation. Calcium homeostasis sensors STIM1 and STIM2 also play an inhibitory role in CACNA1S upregulation upon M . tb infection or Rv2463 stimulation.

    Article Snippet: Rv3416 does not induce CACNA1S on macrophages.

    Techniques: Expressing, Infection

    Calcium homeostasis regulates Rv2463 and M . tb mediated CACNA1S expression on macrophages. J774 cells were treated with inhibitors to internal calcium release (TMB8) or external calcium influx (EGTA) for 1h followed by stimulation with 25 μg/ml Rv2463 (panels A and C) or infection with 2 MOI of M . tb H37Rv (panels B and D) for 48h. CACNA1S expression was monitored by flow cytometry. Grey lines represent cells stimulated in the absence of inhibitors while bold lines represent cells stimulated in the presence of inhibitors. Dotted line represents unstimulated or uninfected cells. One of three independent experiments is shown. P

    Journal: PLoS ONE

    Article Title: Regulation of L-type Voltage Gated Calcium Channel CACNA1S in Macrophages upon Mycobacterium tuberculosis Infection

    doi: 10.1371/journal.pone.0124263

    Figure Lengend Snippet: Calcium homeostasis regulates Rv2463 and M . tb mediated CACNA1S expression on macrophages. J774 cells were treated with inhibitors to internal calcium release (TMB8) or external calcium influx (EGTA) for 1h followed by stimulation with 25 μg/ml Rv2463 (panels A and C) or infection with 2 MOI of M . tb H37Rv (panels B and D) for 48h. CACNA1S expression was monitored by flow cytometry. Grey lines represent cells stimulated in the absence of inhibitors while bold lines represent cells stimulated in the presence of inhibitors. Dotted line represents unstimulated or uninfected cells. One of three independent experiments is shown. P

    Article Snippet: Rv3416 does not induce CACNA1S on macrophages.

    Techniques: Expressing, Infection, Flow Cytometry, Cytometry

    Rv2463 upregulates CACNA1S in a TLR dependent and MyD88 independent pathway on macrophages. J774 cells were transfected with siRNAs against MyD88, TRAF6 or IRAK1 for 36h followed by stimulations with 25 μg/ml Rv2463 or infection with 2 MOI of M . tb H37Rv for 48h. Surface CACNA1S expression was monitored by flow cytometry. Grey lines represent cells transfected with control siRNAs followed by stimulations with Rv2463 or infection with M . tb H37Rv, while bold lines represent cells transfected with specific siRNAs to indicated molecules followed by stimulations with Rv2463 or infection with M . tb H37Rv. Dotted line represents unstimulated or uninfected cells transfected with control siRNAs. One of three independent experiments is shown. P

    Journal: PLoS ONE

    Article Title: Regulation of L-type Voltage Gated Calcium Channel CACNA1S in Macrophages upon Mycobacterium tuberculosis Infection

    doi: 10.1371/journal.pone.0124263

    Figure Lengend Snippet: Rv2463 upregulates CACNA1S in a TLR dependent and MyD88 independent pathway on macrophages. J774 cells were transfected with siRNAs against MyD88, TRAF6 or IRAK1 for 36h followed by stimulations with 25 μg/ml Rv2463 or infection with 2 MOI of M . tb H37Rv for 48h. Surface CACNA1S expression was monitored by flow cytometry. Grey lines represent cells transfected with control siRNAs followed by stimulations with Rv2463 or infection with M . tb H37Rv, while bold lines represent cells transfected with specific siRNAs to indicated molecules followed by stimulations with Rv2463 or infection with M . tb H37Rv. Dotted line represents unstimulated or uninfected cells transfected with control siRNAs. One of three independent experiments is shown. P

    Article Snippet: Rv3416 does not induce CACNA1S on macrophages.

    Techniques: Transfection, Infection, Expressing, Flow Cytometry, Cytometry

    M . tb H37Rv induces the upregulation of CACNA1S on macrophages. J774 macrophages were infected with M . tb H37Rv at indicated Multiplicity of Infection (MOI) for indicated times. CACNA1S expression was monitored by flow cytometry. Bold lines represent infections with M . tb H37Rv while dotted lines represent uninfected controls. Data from one of four independent experiments are shown. P

    Journal: PLoS ONE

    Article Title: Regulation of L-type Voltage Gated Calcium Channel CACNA1S in Macrophages upon Mycobacterium tuberculosis Infection

    doi: 10.1371/journal.pone.0124263

    Figure Lengend Snippet: M . tb H37Rv induces the upregulation of CACNA1S on macrophages. J774 macrophages were infected with M . tb H37Rv at indicated Multiplicity of Infection (MOI) for indicated times. CACNA1S expression was monitored by flow cytometry. Bold lines represent infections with M . tb H37Rv while dotted lines represent uninfected controls. Data from one of four independent experiments are shown. P

    Article Snippet: Rv3416 does not induce CACNA1S on macrophages.

    Techniques: Infection, Expressing, Flow Cytometry, Cytometry

    Rv2463 recruits CREB to CACNA1S promoter in a MyD88 independent and TLR dependent pathway. For Panel A, J774 cells were stimulated with 25 μg/ml Rv2463 for indicated times. Nuclear extracts (NE) were subjected to EMSA with 32 P-labeled oligo (Probe) corresponding to 29 bp in the CACNA1S promoter. For Panel B, nuclear extracts from J774 cells stimulated with 25 μg/ml Rv2463 for 120 min were subjected to EMSA in the presence or absence of indicated concentrations of unlabelled oligo (Cold Probe). For Panel C, J774 cells were transfected with siRNAs to MyD88 or TRAF6 or IRAK1 for 36h followed by stimulations with 25 μg/ml Rv2463 for 120 min. siControl represents cells transfected with control siRNAs. Arrow indicates the specific band. One of three independent experiments is shown. P

    Journal: PLoS ONE

    Article Title: Regulation of L-type Voltage Gated Calcium Channel CACNA1S in Macrophages upon Mycobacterium tuberculosis Infection

    doi: 10.1371/journal.pone.0124263

    Figure Lengend Snippet: Rv2463 recruits CREB to CACNA1S promoter in a MyD88 independent and TLR dependent pathway. For Panel A, J774 cells were stimulated with 25 μg/ml Rv2463 for indicated times. Nuclear extracts (NE) were subjected to EMSA with 32 P-labeled oligo (Probe) corresponding to 29 bp in the CACNA1S promoter. For Panel B, nuclear extracts from J774 cells stimulated with 25 μg/ml Rv2463 for 120 min were subjected to EMSA in the presence or absence of indicated concentrations of unlabelled oligo (Cold Probe). For Panel C, J774 cells were transfected with siRNAs to MyD88 or TRAF6 or IRAK1 for 36h followed by stimulations with 25 μg/ml Rv2463 for 120 min. siControl represents cells transfected with control siRNAs. Arrow indicates the specific band. One of three independent experiments is shown. P

    Article Snippet: Rv3416 does not induce CACNA1S on macrophages.

    Techniques: Labeling, Transfection

    Transcription factors CREB, SOX5 and GATA2 regulate Rv2463 and M . tb mediated CACNA1S expression on macrophages. For Panel A, J774 cells were transfected with siRNAs against CREB or SOX5 or GATA2 for 36h followed by stimulations with 25 μg/ml Rv2463 or infection with 2 MOI of M . tb H37Rv for 48h. CACNA1S expression was monitored by flow cytometry. Grey lines represent cells transfected with control siRNAs followed by stimulations with Rv2463 or infection with M . tb . Bold lines represent cells transfected with specific siRNA to indicated molecules followed by stimulations with Rv2463 or infection with M . tb H37Rv. Dotted lines represent unstimulated cells transfected with control siRNAs. One of three independent experiments is shown. For Panel B, J774 cells were stimulated with 25 μg/ml Rv2463 or infection with 2 MOI of M . tb H37Rv for indicated times and nuclear extracts were western blotted for indicated molecules. Arrow indicates specific band. Numbers below the bands indicates relative intensities of the blots. One of two independent experiments is shown. For Panel A, P

    Journal: PLoS ONE

    Article Title: Regulation of L-type Voltage Gated Calcium Channel CACNA1S in Macrophages upon Mycobacterium tuberculosis Infection

    doi: 10.1371/journal.pone.0124263

    Figure Lengend Snippet: Transcription factors CREB, SOX5 and GATA2 regulate Rv2463 and M . tb mediated CACNA1S expression on macrophages. For Panel A, J774 cells were transfected with siRNAs against CREB or SOX5 or GATA2 for 36h followed by stimulations with 25 μg/ml Rv2463 or infection with 2 MOI of M . tb H37Rv for 48h. CACNA1S expression was monitored by flow cytometry. Grey lines represent cells transfected with control siRNAs followed by stimulations with Rv2463 or infection with M . tb . Bold lines represent cells transfected with specific siRNA to indicated molecules followed by stimulations with Rv2463 or infection with M . tb H37Rv. Dotted lines represent unstimulated cells transfected with control siRNAs. One of three independent experiments is shown. For Panel B, J774 cells were stimulated with 25 μg/ml Rv2463 or infection with 2 MOI of M . tb H37Rv for indicated times and nuclear extracts were western blotted for indicated molecules. Arrow indicates specific band. Numbers below the bands indicates relative intensities of the blots. One of two independent experiments is shown. For Panel A, P

    Article Snippet: Rv3416 does not induce CACNA1S on macrophages.

    Techniques: Expressing, Transfection, Infection, Flow Cytometry, Cytometry, Western Blot

    Cross regulation of CACNA1S, ROS and pCREB during Rv2463 stimulation or M . tb infection. For Panel A, J774 cells were stimulated with 25 μg/ml Rv2463 or infected with 2 MOI M . tb H37Rv for 48h in the presence (bold lines) or absence (dotted lines) of indicated inhibitors to ROS. CACNA1S levels were monitored by flow cytometry. For Panel B, J774 cells were transfected with siRNAs to CACNA1S for 36h followed by stimulation with 25 μg/ml Rv2463 for 48h. ROS levels were monitored by FACS. Grey line represents Rv2463 stimulated cells transfected with control siRNAs, while bold line represents Rv2463 stimulated cells transfected with siRNAs specific to CACNA1S. Dotted line represents unstimulated cells transfected with control siRNAs. For Panel C, J774 cells were stimulated with 25 μg/ml Rv2463 or infected with 2 MOI M . tb H37Rv for indicated times in the presence or absence of indicated inhibitors to ROS and nuclear extracts were probed for pCREB levels. Numbers below the blots indicate relative intensities of the bands. Arrow shows the specific band. Data from one of three experiments is shown. In Panel A, P

    Journal: PLoS ONE

    Article Title: Regulation of L-type Voltage Gated Calcium Channel CACNA1S in Macrophages upon Mycobacterium tuberculosis Infection

    doi: 10.1371/journal.pone.0124263

    Figure Lengend Snippet: Cross regulation of CACNA1S, ROS and pCREB during Rv2463 stimulation or M . tb infection. For Panel A, J774 cells were stimulated with 25 μg/ml Rv2463 or infected with 2 MOI M . tb H37Rv for 48h in the presence (bold lines) or absence (dotted lines) of indicated inhibitors to ROS. CACNA1S levels were monitored by flow cytometry. For Panel B, J774 cells were transfected with siRNAs to CACNA1S for 36h followed by stimulation with 25 μg/ml Rv2463 for 48h. ROS levels were monitored by FACS. Grey line represents Rv2463 stimulated cells transfected with control siRNAs, while bold line represents Rv2463 stimulated cells transfected with siRNAs specific to CACNA1S. Dotted line represents unstimulated cells transfected with control siRNAs. For Panel C, J774 cells were stimulated with 25 μg/ml Rv2463 or infected with 2 MOI M . tb H37Rv for indicated times in the presence or absence of indicated inhibitors to ROS and nuclear extracts were probed for pCREB levels. Numbers below the blots indicate relative intensities of the bands. Arrow shows the specific band. Data from one of three experiments is shown. In Panel A, P

    Article Snippet: Rv3416 does not induce CACNA1S on macrophages.

    Techniques: Infection, Flow Cytometry, Cytometry, Transfection, FACS

    Molecular sensors of calcium homeostasis positively regulate Rv2463 mediated CACNA1S expression on macrophages. J774 cells were transfected with siRNAs against ORAI1, STIM1 or STIM2 for 36h followed by stimulations with 25 μg/ml Rv2463 or infection with 2 MOI of M . tb H37Rv for 48h. CACNA1S expression was monitored by flow cytometry. Grey lines represent cells transfected with control siRNAs followed by stimulations with Rv2463 or infection with M . tb . Bold lines represent cells transfected with specific siRNA to indicated molecules followed by stimulations with Rv2463 or infection with M . tb . Dotted lines represent unstimulated or uninfected cells transfected with control siRNAs. One of three independent experiments is shown. P

    Journal: PLoS ONE

    Article Title: Regulation of L-type Voltage Gated Calcium Channel CACNA1S in Macrophages upon Mycobacterium tuberculosis Infection

    doi: 10.1371/journal.pone.0124263

    Figure Lengend Snippet: Molecular sensors of calcium homeostasis positively regulate Rv2463 mediated CACNA1S expression on macrophages. J774 cells were transfected with siRNAs against ORAI1, STIM1 or STIM2 for 36h followed by stimulations with 25 μg/ml Rv2463 or infection with 2 MOI of M . tb H37Rv for 48h. CACNA1S expression was monitored by flow cytometry. Grey lines represent cells transfected with control siRNAs followed by stimulations with Rv2463 or infection with M . tb . Bold lines represent cells transfected with specific siRNA to indicated molecules followed by stimulations with Rv2463 or infection with M . tb . Dotted lines represent unstimulated or uninfected cells transfected with control siRNAs. One of three independent experiments is shown. P

    Article Snippet: Rv3416 does not induce CACNA1S on macrophages.

    Techniques: Expressing, Transfection, Infection, Flow Cytometry, Cytometry

    Rv2463 induces the upregulation of CACNA1S on macrophages. J774 macrophages were stimulated with Rv2463 at indicated concentrations and for indicated times. CACNA1S expression on cell surface was monitored by flow cytometry. Bold lines represent stimulations with Rv2463 while dotted lines represent unstimulated controls. Data from one of four independent experiments are shown. P

    Journal: PLoS ONE

    Article Title: Regulation of L-type Voltage Gated Calcium Channel CACNA1S in Macrophages upon Mycobacterium tuberculosis Infection

    doi: 10.1371/journal.pone.0124263

    Figure Lengend Snippet: Rv2463 induces the upregulation of CACNA1S on macrophages. J774 macrophages were stimulated with Rv2463 at indicated concentrations and for indicated times. CACNA1S expression on cell surface was monitored by flow cytometry. Bold lines represent stimulations with Rv2463 while dotted lines represent unstimulated controls. Data from one of four independent experiments are shown. P

    Article Snippet: Rv3416 does not induce CACNA1S on macrophages.

    Techniques: Expressing, Flow Cytometry, Cytometry

    Confocal images of CACNA1S following stimulations with Rv2463 and M . tb on macrophages. J774 cells were stimulated with 25 μg/ml Rv2463 or 2 MOI M . tb H37Rv for 48h. CACNA1S expression was monitored by confocal imaging ( see Materials and Methods ). Blue colour represents staining of nucleus with DAPI while green colour represents CACNA1S staining with streptavidin conjugated FITC bound to biotinylated antibody to CACNA1S. A representative image of 10 fields is shown. P

    Journal: PLoS ONE

    Article Title: Regulation of L-type Voltage Gated Calcium Channel CACNA1S in Macrophages upon Mycobacterium tuberculosis Infection

    doi: 10.1371/journal.pone.0124263

    Figure Lengend Snippet: Confocal images of CACNA1S following stimulations with Rv2463 and M . tb on macrophages. J774 cells were stimulated with 25 μg/ml Rv2463 or 2 MOI M . tb H37Rv for 48h. CACNA1S expression was monitored by confocal imaging ( see Materials and Methods ). Blue colour represents staining of nucleus with DAPI while green colour represents CACNA1S staining with streptavidin conjugated FITC bound to biotinylated antibody to CACNA1S. A representative image of 10 fields is shown. P

    Article Snippet: Rv3416 does not induce CACNA1S on macrophages.

    Techniques: Expressing, Imaging, Staining