Structured Review

Cayman Chemical zd7288
Functional properties of oligodendrocyte HCN channels. A , Whole-cell voltage-clamp currents recorded from an MBP-positive oligodendrocyte in response to an activation protocol (detailed in the bottom panel) in the absence (top) and presence (middle, blue) of HCN channel blocker <t>ZD7288</t> (30 μ m ). Bottom, Leak-subtracted currents from the example. HCN currents were measured in the presence of Ba 2+ (1 m m ) in order to block the influence of inwardly rectifying potassium channel currents (see Materials and Methods for full details of the protocol). B , Whole-cell voltage-clamp currents recorded from an MBP-negative oligodendrocyte progenitor in response to an activation protocol (detailed in the bottom panel) in the absence (black) and presence (blue) of HCN channel blocker ZD7288 (30 μ m ). C , The black plot represents the mean ± SE activation curve of ZD7288-sensitive tail currents ( n = 6). The mean RMP of oligodendrocytes (dashed line) predicts ∼15% channel opening. The plot in red represents the mean ± SE activation curve of ZD7288-sensitive tail currents in the presence of 8-bromo-cAMP [8-Br-cAMP] ( n = 6). D , Current-clamp recording from an MBP-positive oligodendrocyte where ZD7288 application generates hyperpolarization of the RMP.
Zd7288, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/zd7288/product/Cayman Chemical
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
zd7288 - by Bioz Stars, 2023-02
93/100 stars

Images

1) Product Images from "Oligodendrocyte HCN2 Channels Regulate Myelin Sheath Length"

Article Title: Oligodendrocyte HCN2 Channels Regulate Myelin Sheath Length

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.2463-20.2021

Functional properties of oligodendrocyte HCN channels. A , Whole-cell voltage-clamp currents recorded from an MBP-positive oligodendrocyte in response to an activation protocol (detailed in the bottom panel) in the absence (top) and presence (middle, blue) of HCN channel blocker ZD7288 (30 μ m ). Bottom, Leak-subtracted currents from the example. HCN currents were measured in the presence of Ba 2+ (1 m m ) in order to block the influence of inwardly rectifying potassium channel currents (see Materials and Methods for full details of the protocol). B , Whole-cell voltage-clamp currents recorded from an MBP-negative oligodendrocyte progenitor in response to an activation protocol (detailed in the bottom panel) in the absence (black) and presence (blue) of HCN channel blocker ZD7288 (30 μ m ). C , The black plot represents the mean ± SE activation curve of ZD7288-sensitive tail currents ( n = 6). The mean RMP of oligodendrocytes (dashed line) predicts ∼15% channel opening. The plot in red represents the mean ± SE activation curve of ZD7288-sensitive tail currents in the presence of 8-bromo-cAMP [8-Br-cAMP] ( n = 6). D , Current-clamp recording from an MBP-positive oligodendrocyte where ZD7288 application generates hyperpolarization of the RMP.
Figure Legend Snippet: Functional properties of oligodendrocyte HCN channels. A , Whole-cell voltage-clamp currents recorded from an MBP-positive oligodendrocyte in response to an activation protocol (detailed in the bottom panel) in the absence (top) and presence (middle, blue) of HCN channel blocker ZD7288 (30 μ m ). Bottom, Leak-subtracted currents from the example. HCN currents were measured in the presence of Ba 2+ (1 m m ) in order to block the influence of inwardly rectifying potassium channel currents (see Materials and Methods for full details of the protocol). B , Whole-cell voltage-clamp currents recorded from an MBP-negative oligodendrocyte progenitor in response to an activation protocol (detailed in the bottom panel) in the absence (black) and presence (blue) of HCN channel blocker ZD7288 (30 μ m ). C , The black plot represents the mean ± SE activation curve of ZD7288-sensitive tail currents ( n = 6). The mean RMP of oligodendrocytes (dashed line) predicts ∼15% channel opening. The plot in red represents the mean ± SE activation curve of ZD7288-sensitive tail currents in the presence of 8-bromo-cAMP [8-Br-cAMP] ( n = 6). D , Current-clamp recording from an MBP-positive oligodendrocyte where ZD7288 application generates hyperpolarization of the RMP.

Techniques Used: Functional Assay, Activation Assay, Blocking Assay

HCN channels regulate the length of myelin sheaths formed in oligodendrocyte-purified cultures. A , Representative MBP-positive oligodendrocytes after 14 d cultured on electrospun microfibers in the presence of either 0.1% DMSO or 10 μ m ZD7288. Scale bars, 10 µm. B , Mean ± SE percentage of MBP-positive cells on microfibers: DMSO, 60.77 ± 2.972%; ZD7288, 61.76 ± 2.659%; n = 4 independent cultures; p = 0.8857, Mann–Whitney test. C , Mean myelin sheath length generated on microfibers. Values are reported as the mean ± SE; DMSO: 19.03 ± 0.6101µm; n = 342 sheaths from five independent cultures; ZD7288: 14.23 ± 0.9092 µm; n = 241 sheaths from four independent cultures; p = 0.0159, Mann–Whitney test. D , Histogram representation of the frequency of sheath lengths generated by oligodendrocytes as assessed by measuring complete MBP-positive sheaths surrounding microfibers in 5 µm bins. Pooled data from n = 4–5 independent cultures. E , Pooled data of the number of complete sheaths formed by individual oligodendrocytes in microfiber cultures. Values are reported as the mean ± SD; DMSO: 3.613 ± 2.4; n = 93 cells; ZD7288: 3.280 ± 1.935; n = 75 cells; pooled data from n = 4–5 independent cultures; p = 0.5695, Mann–Whitney test. F , qPCR of two-dimensional 3 DIV rat oligodendroglial cultures treated with ZD7288 for the expression of Mbp normalized to Gapdh. DMSO, 1.003 ± 0.03327; ZD7288, 0.9658 ± 0.03675; n = 4 independent cultures; p = 0.4857, Mann–Whitney test. G , Representative Western blot images for loading control β-actin, MBP, and CNPase from two-dimensional rat oligodendroglial cultures treated with ZD7288. H , MBP protein expression in arbitrary units (A.U.) normalized to β-actin levels. Values are reported as the mean ± SD; DMSO, 12.12 ± 2.084; ZD7288, 7.558 ± 2.540; n = 4 independent cultures; p = 0.0286, Mann–Whitney test.
Figure Legend Snippet: HCN channels regulate the length of myelin sheaths formed in oligodendrocyte-purified cultures. A , Representative MBP-positive oligodendrocytes after 14 d cultured on electrospun microfibers in the presence of either 0.1% DMSO or 10 μ m ZD7288. Scale bars, 10 µm. B , Mean ± SE percentage of MBP-positive cells on microfibers: DMSO, 60.77 ± 2.972%; ZD7288, 61.76 ± 2.659%; n = 4 independent cultures; p = 0.8857, Mann–Whitney test. C , Mean myelin sheath length generated on microfibers. Values are reported as the mean ± SE; DMSO: 19.03 ± 0.6101µm; n = 342 sheaths from five independent cultures; ZD7288: 14.23 ± 0.9092 µm; n = 241 sheaths from four independent cultures; p = 0.0159, Mann–Whitney test. D , Histogram representation of the frequency of sheath lengths generated by oligodendrocytes as assessed by measuring complete MBP-positive sheaths surrounding microfibers in 5 µm bins. Pooled data from n = 4–5 independent cultures. E , Pooled data of the number of complete sheaths formed by individual oligodendrocytes in microfiber cultures. Values are reported as the mean ± SD; DMSO: 3.613 ± 2.4; n = 93 cells; ZD7288: 3.280 ± 1.935; n = 75 cells; pooled data from n = 4–5 independent cultures; p = 0.5695, Mann–Whitney test. F , qPCR of two-dimensional 3 DIV rat oligodendroglial cultures treated with ZD7288 for the expression of Mbp normalized to Gapdh. DMSO, 1.003 ± 0.03327; ZD7288, 0.9658 ± 0.03675; n = 4 independent cultures; p = 0.4857, Mann–Whitney test. G , Representative Western blot images for loading control β-actin, MBP, and CNPase from two-dimensional rat oligodendroglial cultures treated with ZD7288. H , MBP protein expression in arbitrary units (A.U.) normalized to β-actin levels. Values are reported as the mean ± SD; DMSO, 12.12 ± 2.084; ZD7288, 7.558 ± 2.540; n = 4 independent cultures; p = 0.0286, Mann–Whitney test.

Techniques Used: Purification, Cell Culture, MANN-WHITNEY, Generated, Expressing, Western Blot


Structured Review

Cayman Chemical zd 7288
( A ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated sodium channel blocker tetrodotoxin (TTX; 1 µM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( B ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated potassium channel blocker tetraethylammonium (TEA; 10 mM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( C ) Forskolin-mediated reactivation in the presence of the HCN channel blockers <t>ZD</t> <t>7288</t> (10μM) quantified as the numbers of Us11-GFP-positive neurons at 3 days post-reactivation. ( D ) The effect of ZD 7288 on the HSV lytic gene transcript ICP27 during Phase I reactivation measured at 20 hr post-forskolin treatment by RT-qPCR. In A-D individual experimental replicates are represented along with the mean and SEM. ( E and F ) Quantification of the relative nuclear staining for H3K9me3/S10p and γH2AX in SCG neurons at 5 hr post-forskolin treatment and in the presence of ZD 7288 from >800 cells/condition from two independent experiments. Data are plotted around the mean, with the boxes representing the 25 th -75 th percentiles and the whiskers the 5 st -95 th percentiles. Statistical comparisons were made using a one-way ANOVA with a Tukey’s multiple comparison ( A–D ) or two-tailed unpaired t-test ( E–F ). *p<0.05, **p<0.01, ***p<0.001. In A-D individual experimental replicates are represented. Figure 4—source data 1. Quantification of GFP-positive neurons, RT-qPCR and nuclear staining intensity for .
Zd 7288, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/zd 7288/product/Cayman Chemical
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
zd 7288 - by Bioz Stars, 2023-02
93/100 stars

Images

1) Product Images from "Neuronal hyperexcitability is a DLK-dependent trigger of herpes simplex virus reactivation that can be induced by IL-1"

Article Title: Neuronal hyperexcitability is a DLK-dependent trigger of herpes simplex virus reactivation that can be induced by IL-1

Journal: eLife

doi: 10.7554/eLife.58037

( A ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated sodium channel blocker tetrodotoxin (TTX; 1 µM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( B ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated potassium channel blocker tetraethylammonium (TEA; 10 mM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( C ) Forskolin-mediated reactivation in the presence of the HCN channel blockers ZD 7288 (10μM) quantified as the numbers of Us11-GFP-positive neurons at 3 days post-reactivation. ( D ) The effect of ZD 7288 on the HSV lytic gene transcript ICP27 during Phase I reactivation measured at 20 hr post-forskolin treatment by RT-qPCR. In A-D individual experimental replicates are represented along with the mean and SEM. ( E and F ) Quantification of the relative nuclear staining for H3K9me3/S10p and γH2AX in SCG neurons at 5 hr post-forskolin treatment and in the presence of ZD 7288 from >800 cells/condition from two independent experiments. Data are plotted around the mean, with the boxes representing the 25 th -75 th percentiles and the whiskers the 5 st -95 th percentiles. Statistical comparisons were made using a one-way ANOVA with a Tukey’s multiple comparison ( A–D ) or two-tailed unpaired t-test ( E–F ). *p<0.05, **p<0.01, ***p<0.001. In A-D individual experimental replicates are represented. Figure 4—source data 1. Quantification of GFP-positive neurons, RT-qPCR and nuclear staining intensity for .
Figure Legend Snippet: ( A ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated sodium channel blocker tetrodotoxin (TTX; 1 µM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( B ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated potassium channel blocker tetraethylammonium (TEA; 10 mM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( C ) Forskolin-mediated reactivation in the presence of the HCN channel blockers ZD 7288 (10μM) quantified as the numbers of Us11-GFP-positive neurons at 3 days post-reactivation. ( D ) The effect of ZD 7288 on the HSV lytic gene transcript ICP27 during Phase I reactivation measured at 20 hr post-forskolin treatment by RT-qPCR. In A-D individual experimental replicates are represented along with the mean and SEM. ( E and F ) Quantification of the relative nuclear staining for H3K9me3/S10p and γH2AX in SCG neurons at 5 hr post-forskolin treatment and in the presence of ZD 7288 from >800 cells/condition from two independent experiments. Data are plotted around the mean, with the boxes representing the 25 th -75 th percentiles and the whiskers the 5 st -95 th percentiles. Statistical comparisons were made using a one-way ANOVA with a Tukey’s multiple comparison ( A–D ) or two-tailed unpaired t-test ( E–F ). *p<0.05, **p<0.01, ***p<0.001. In A-D individual experimental replicates are represented. Figure 4—source data 1. Quantification of GFP-positive neurons, RT-qPCR and nuclear staining intensity for .

Techniques Used: Infection, Quantitative RT-PCR, Staining, Two Tailed Test


Figure Legend Snippet:

Techniques Used: Recombinant, Plasmid Preparation, Blocking Assay, Sequencing, shRNA, SYBR Green Assay, Staining


Structured Review

Cayman Chemical zd 7288
(A) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated sodium channel blocker tetrodotoxin (TTX; 1μM) and the number of Us11-GFP positive neurons quantified at 3 days post-reactivation. (B) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated potassium channel blocker tetraethylammonium (TEA; 10 mM) and the number of Us11-GFP positive neurons quantified at 3 days post-reactivation. (C) Forskolin-mediated reactivation in the presence of the HCN channel blockers <t>ZD</t> <t>7288</t> (10μM) quantified as the numbers of Us11-GFP positive neurons at 3 days post-reactivation. (D) The effect of ZD 7288 on the HSV lytic gene transcript ICP27 during Phase I reactivation measured at 20h post-forskolin treatment by RT-qPCR. Individual experimental replicates are represented. (E and F) Quantification of the relative nuclear staining for H3K9me3/S10p and γH2AX in SCG neurons at 5h post-forskolin treatment and in the presence of ZD 7288 from two independent experiments. Statistical comparisons were made using a one-way ANOVA with a Tukey’s multiple comparison (A-D) or two-tailed unpaired t-test (E-F). *P<0.05, ** P<0.01, ***P<0.001.
Zd 7288, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/zd 7288/product/Cayman Chemical
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
zd 7288 - by Bioz Stars, 2023-02
93/100 stars

Images

1) Product Images from "Neuronal hyperexcitability is a DLK-dependent trigger of HSV-1 reactivation that can be induced by IL-1"

Article Title: Neuronal hyperexcitability is a DLK-dependent trigger of HSV-1 reactivation that can be induced by IL-1

Journal: bioRxiv

doi: 10.1101/2020.04.16.044875

(A) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated sodium channel blocker tetrodotoxin (TTX; 1μM) and the number of Us11-GFP positive neurons quantified at 3 days post-reactivation. (B) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated potassium channel blocker tetraethylammonium (TEA; 10 mM) and the number of Us11-GFP positive neurons quantified at 3 days post-reactivation. (C) Forskolin-mediated reactivation in the presence of the HCN channel blockers ZD 7288 (10μM) quantified as the numbers of Us11-GFP positive neurons at 3 days post-reactivation. (D) The effect of ZD 7288 on the HSV lytic gene transcript ICP27 during Phase I reactivation measured at 20h post-forskolin treatment by RT-qPCR. Individual experimental replicates are represented. (E and F) Quantification of the relative nuclear staining for H3K9me3/S10p and γH2AX in SCG neurons at 5h post-forskolin treatment and in the presence of ZD 7288 from two independent experiments. Statistical comparisons were made using a one-way ANOVA with a Tukey’s multiple comparison (A-D) or two-tailed unpaired t-test (E-F). *P<0.05, ** P<0.01, ***P<0.001.
Figure Legend Snippet: (A) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated sodium channel blocker tetrodotoxin (TTX; 1μM) and the number of Us11-GFP positive neurons quantified at 3 days post-reactivation. (B) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated potassium channel blocker tetraethylammonium (TEA; 10 mM) and the number of Us11-GFP positive neurons quantified at 3 days post-reactivation. (C) Forskolin-mediated reactivation in the presence of the HCN channel blockers ZD 7288 (10μM) quantified as the numbers of Us11-GFP positive neurons at 3 days post-reactivation. (D) The effect of ZD 7288 on the HSV lytic gene transcript ICP27 during Phase I reactivation measured at 20h post-forskolin treatment by RT-qPCR. Individual experimental replicates are represented. (E and F) Quantification of the relative nuclear staining for H3K9me3/S10p and γH2AX in SCG neurons at 5h post-forskolin treatment and in the presence of ZD 7288 from two independent experiments. Statistical comparisons were made using a one-way ANOVA with a Tukey’s multiple comparison (A-D) or two-tailed unpaired t-test (E-F). *P<0.05, ** P<0.01, ***P<0.001.

Techniques Used: Infection, Quantitative RT-PCR, Staining, Two Tailed Test

(A and B) Latently infected cultures were reactivated with forskolin in the presence of the HCN channel inhibitors ivabradine (20μM; A) and CsCl (3mM; B). Latently infected cultures were reactivated with forskolin in the presence of the HCN inhibitor ZD 7288 (10 μM) and viral lytic transcripts measured at 20h post-reactivation (C and D). Individual experimental replicates are represented. Statistical comparisons were made using a one-way ANOVA with a Tukey’s multiple comparison. *P<0.05, ** P<0.01.
Figure Legend Snippet: (A and B) Latently infected cultures were reactivated with forskolin in the presence of the HCN channel inhibitors ivabradine (20μM; A) and CsCl (3mM; B). Latently infected cultures were reactivated with forskolin in the presence of the HCN inhibitor ZD 7288 (10 μM) and viral lytic transcripts measured at 20h post-reactivation (C and D). Individual experimental replicates are represented. Statistical comparisons were made using a one-way ANOVA with a Tukey’s multiple comparison. *P<0.05, ** P<0.01.

Techniques Used: Infection

(A) Adult P28 SCG neurons were treated with IL-1β (30ng/mL) for 15 hrs and stained for H3K9me3/S10p, γH2AX and beta II-tubulin to mark neurons. (B-C) Quantification of the intensity of H3K9me3/S10p and γH2AX in neuronal nuclei following forskolin treatment from two independent experiments. (D). Addition of IL-1β to latently infected cultures of mature SCG neurons triggers HSV reactivation. (E). Quantification of IL-1β induced reactivation in the presence of the voltage gated sodium channel blocker TTX (1μM), the HCN channel blocker ZD 7288 (10μM) and the DLK inhibitor GNE-3511 (4μM). In D and E individual experimental replicates are represented. Statistical comparisons were made using or two-tailed unpaired t-test (D) or a one-way ANOVA with a Tukey’s multiple comparison (B,C & E). *P<0.05, **P<0.01, *** P<0.001.
Figure Legend Snippet: (A) Adult P28 SCG neurons were treated with IL-1β (30ng/mL) for 15 hrs and stained for H3K9me3/S10p, γH2AX and beta II-tubulin to mark neurons. (B-C) Quantification of the intensity of H3K9me3/S10p and γH2AX in neuronal nuclei following forskolin treatment from two independent experiments. (D). Addition of IL-1β to latently infected cultures of mature SCG neurons triggers HSV reactivation. (E). Quantification of IL-1β induced reactivation in the presence of the voltage gated sodium channel blocker TTX (1μM), the HCN channel blocker ZD 7288 (10μM) and the DLK inhibitor GNE-3511 (4μM). In D and E individual experimental replicates are represented. Statistical comparisons were made using or two-tailed unpaired t-test (D) or a one-way ANOVA with a Tukey’s multiple comparison (B,C & E). *P<0.05, **P<0.01, *** P<0.001.

Techniques Used: Staining, Infection, Two Tailed Test

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    Cayman Chemical zd7288
    Functional properties of oligodendrocyte HCN channels. A , Whole-cell voltage-clamp currents recorded from an MBP-positive oligodendrocyte in response to an activation protocol (detailed in the bottom panel) in the absence (top) and presence (middle, blue) of HCN channel blocker <t>ZD7288</t> (30 μ m ). Bottom, Leak-subtracted currents from the example. HCN currents were measured in the presence of Ba 2+ (1 m m ) in order to block the influence of inwardly rectifying potassium channel currents (see Materials and Methods for full details of the protocol). B , Whole-cell voltage-clamp currents recorded from an MBP-negative oligodendrocyte progenitor in response to an activation protocol (detailed in the bottom panel) in the absence (black) and presence (blue) of HCN channel blocker ZD7288 (30 μ m ). C , The black plot represents the mean ± SE activation curve of ZD7288-sensitive tail currents ( n = 6). The mean RMP of oligodendrocytes (dashed line) predicts ∼15% channel opening. The plot in red represents the mean ± SE activation curve of ZD7288-sensitive tail currents in the presence of 8-bromo-cAMP [8-Br-cAMP] ( n = 6). D , Current-clamp recording from an MBP-positive oligodendrocyte where ZD7288 application generates hyperpolarization of the RMP.
    Zd7288, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/zd7288/product/Cayman Chemical
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    zd7288 - by Bioz Stars, 2023-02
    93/100 stars
      Buy from Supplier

    93
    Cayman Chemical zd 7288
    ( A ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated sodium channel blocker tetrodotoxin (TTX; 1 µM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( B ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated potassium channel blocker tetraethylammonium (TEA; 10 mM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( C ) Forskolin-mediated reactivation in the presence of the HCN channel blockers <t>ZD</t> <t>7288</t> (10μM) quantified as the numbers of Us11-GFP-positive neurons at 3 days post-reactivation. ( D ) The effect of ZD 7288 on the HSV lytic gene transcript ICP27 during Phase I reactivation measured at 20 hr post-forskolin treatment by RT-qPCR. In A-D individual experimental replicates are represented along with the mean and SEM. ( E and F ) Quantification of the relative nuclear staining for H3K9me3/S10p and γH2AX in SCG neurons at 5 hr post-forskolin treatment and in the presence of ZD 7288 from >800 cells/condition from two independent experiments. Data are plotted around the mean, with the boxes representing the 25 th -75 th percentiles and the whiskers the 5 st -95 th percentiles. Statistical comparisons were made using a one-way ANOVA with a Tukey’s multiple comparison ( A–D ) or two-tailed unpaired t-test ( E–F ). *p<0.05, **p<0.01, ***p<0.001. In A-D individual experimental replicates are represented. Figure 4—source data 1. Quantification of GFP-positive neurons, RT-qPCR and nuclear staining intensity for .
    Zd 7288, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/zd 7288/product/Cayman Chemical
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    zd 7288 - by Bioz Stars, 2023-02
    93/100 stars
      Buy from Supplier

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    Functional properties of oligodendrocyte HCN channels. A , Whole-cell voltage-clamp currents recorded from an MBP-positive oligodendrocyte in response to an activation protocol (detailed in the bottom panel) in the absence (top) and presence (middle, blue) of HCN channel blocker ZD7288 (30 μ m ). Bottom, Leak-subtracted currents from the example. HCN currents were measured in the presence of Ba 2+ (1 m m ) in order to block the influence of inwardly rectifying potassium channel currents (see Materials and Methods for full details of the protocol). B , Whole-cell voltage-clamp currents recorded from an MBP-negative oligodendrocyte progenitor in response to an activation protocol (detailed in the bottom panel) in the absence (black) and presence (blue) of HCN channel blocker ZD7288 (30 μ m ). C , The black plot represents the mean ± SE activation curve of ZD7288-sensitive tail currents ( n = 6). The mean RMP of oligodendrocytes (dashed line) predicts ∼15% channel opening. The plot in red represents the mean ± SE activation curve of ZD7288-sensitive tail currents in the presence of 8-bromo-cAMP [8-Br-cAMP] ( n = 6). D , Current-clamp recording from an MBP-positive oligodendrocyte where ZD7288 application generates hyperpolarization of the RMP.

    Journal: The Journal of Neuroscience

    Article Title: Oligodendrocyte HCN2 Channels Regulate Myelin Sheath Length

    doi: 10.1523/JNEUROSCI.2463-20.2021

    Figure Lengend Snippet: Functional properties of oligodendrocyte HCN channels. A , Whole-cell voltage-clamp currents recorded from an MBP-positive oligodendrocyte in response to an activation protocol (detailed in the bottom panel) in the absence (top) and presence (middle, blue) of HCN channel blocker ZD7288 (30 μ m ). Bottom, Leak-subtracted currents from the example. HCN currents were measured in the presence of Ba 2+ (1 m m ) in order to block the influence of inwardly rectifying potassium channel currents (see Materials and Methods for full details of the protocol). B , Whole-cell voltage-clamp currents recorded from an MBP-negative oligodendrocyte progenitor in response to an activation protocol (detailed in the bottom panel) in the absence (black) and presence (blue) of HCN channel blocker ZD7288 (30 μ m ). C , The black plot represents the mean ± SE activation curve of ZD7288-sensitive tail currents ( n = 6). The mean RMP of oligodendrocytes (dashed line) predicts ∼15% channel opening. The plot in red represents the mean ± SE activation curve of ZD7288-sensitive tail currents in the presence of 8-bromo-cAMP [8-Br-cAMP] ( n = 6). D , Current-clamp recording from an MBP-positive oligodendrocyte where ZD7288 application generates hyperpolarization of the RMP.

    Article Snippet: In some experiments, ZD7288 (catalog #15228, Cayman Chemical) was added to block HCN channel function, as described further below.

    Techniques: Functional Assay, Activation Assay, Blocking Assay

    HCN channels regulate the length of myelin sheaths formed in oligodendrocyte-purified cultures. A , Representative MBP-positive oligodendrocytes after 14 d cultured on electrospun microfibers in the presence of either 0.1% DMSO or 10 μ m ZD7288. Scale bars, 10 µm. B , Mean ± SE percentage of MBP-positive cells on microfibers: DMSO, 60.77 ± 2.972%; ZD7288, 61.76 ± 2.659%; n = 4 independent cultures; p = 0.8857, Mann–Whitney test. C , Mean myelin sheath length generated on microfibers. Values are reported as the mean ± SE; DMSO: 19.03 ± 0.6101µm; n = 342 sheaths from five independent cultures; ZD7288: 14.23 ± 0.9092 µm; n = 241 sheaths from four independent cultures; p = 0.0159, Mann–Whitney test. D , Histogram representation of the frequency of sheath lengths generated by oligodendrocytes as assessed by measuring complete MBP-positive sheaths surrounding microfibers in 5 µm bins. Pooled data from n = 4–5 independent cultures. E , Pooled data of the number of complete sheaths formed by individual oligodendrocytes in microfiber cultures. Values are reported as the mean ± SD; DMSO: 3.613 ± 2.4; n = 93 cells; ZD7288: 3.280 ± 1.935; n = 75 cells; pooled data from n = 4–5 independent cultures; p = 0.5695, Mann–Whitney test. F , qPCR of two-dimensional 3 DIV rat oligodendroglial cultures treated with ZD7288 for the expression of Mbp normalized to Gapdh. DMSO, 1.003 ± 0.03327; ZD7288, 0.9658 ± 0.03675; n = 4 independent cultures; p = 0.4857, Mann–Whitney test. G , Representative Western blot images for loading control β-actin, MBP, and CNPase from two-dimensional rat oligodendroglial cultures treated with ZD7288. H , MBP protein expression in arbitrary units (A.U.) normalized to β-actin levels. Values are reported as the mean ± SD; DMSO, 12.12 ± 2.084; ZD7288, 7.558 ± 2.540; n = 4 independent cultures; p = 0.0286, Mann–Whitney test.

    Journal: The Journal of Neuroscience

    Article Title: Oligodendrocyte HCN2 Channels Regulate Myelin Sheath Length

    doi: 10.1523/JNEUROSCI.2463-20.2021

    Figure Lengend Snippet: HCN channels regulate the length of myelin sheaths formed in oligodendrocyte-purified cultures. A , Representative MBP-positive oligodendrocytes after 14 d cultured on electrospun microfibers in the presence of either 0.1% DMSO or 10 μ m ZD7288. Scale bars, 10 µm. B , Mean ± SE percentage of MBP-positive cells on microfibers: DMSO, 60.77 ± 2.972%; ZD7288, 61.76 ± 2.659%; n = 4 independent cultures; p = 0.8857, Mann–Whitney test. C , Mean myelin sheath length generated on microfibers. Values are reported as the mean ± SE; DMSO: 19.03 ± 0.6101µm; n = 342 sheaths from five independent cultures; ZD7288: 14.23 ± 0.9092 µm; n = 241 sheaths from four independent cultures; p = 0.0159, Mann–Whitney test. D , Histogram representation of the frequency of sheath lengths generated by oligodendrocytes as assessed by measuring complete MBP-positive sheaths surrounding microfibers in 5 µm bins. Pooled data from n = 4–5 independent cultures. E , Pooled data of the number of complete sheaths formed by individual oligodendrocytes in microfiber cultures. Values are reported as the mean ± SD; DMSO: 3.613 ± 2.4; n = 93 cells; ZD7288: 3.280 ± 1.935; n = 75 cells; pooled data from n = 4–5 independent cultures; p = 0.5695, Mann–Whitney test. F , qPCR of two-dimensional 3 DIV rat oligodendroglial cultures treated with ZD7288 for the expression of Mbp normalized to Gapdh. DMSO, 1.003 ± 0.03327; ZD7288, 0.9658 ± 0.03675; n = 4 independent cultures; p = 0.4857, Mann–Whitney test. G , Representative Western blot images for loading control β-actin, MBP, and CNPase from two-dimensional rat oligodendroglial cultures treated with ZD7288. H , MBP protein expression in arbitrary units (A.U.) normalized to β-actin levels. Values are reported as the mean ± SD; DMSO, 12.12 ± 2.084; ZD7288, 7.558 ± 2.540; n = 4 independent cultures; p = 0.0286, Mann–Whitney test.

    Article Snippet: In some experiments, ZD7288 (catalog #15228, Cayman Chemical) was added to block HCN channel function, as described further below.

    Techniques: Purification, Cell Culture, MANN-WHITNEY, Generated, Expressing, Western Blot

    ( A ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated sodium channel blocker tetrodotoxin (TTX; 1 µM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( B ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated potassium channel blocker tetraethylammonium (TEA; 10 mM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( C ) Forskolin-mediated reactivation in the presence of the HCN channel blockers ZD 7288 (10μM) quantified as the numbers of Us11-GFP-positive neurons at 3 days post-reactivation. ( D ) The effect of ZD 7288 on the HSV lytic gene transcript ICP27 during Phase I reactivation measured at 20 hr post-forskolin treatment by RT-qPCR. In A-D individual experimental replicates are represented along with the mean and SEM. ( E and F ) Quantification of the relative nuclear staining for H3K9me3/S10p and γH2AX in SCG neurons at 5 hr post-forskolin treatment and in the presence of ZD 7288 from >800 cells/condition from two independent experiments. Data are plotted around the mean, with the boxes representing the 25 th -75 th percentiles and the whiskers the 5 st -95 th percentiles. Statistical comparisons were made using a one-way ANOVA with a Tukey’s multiple comparison ( A–D ) or two-tailed unpaired t-test ( E–F ). *p<0.05, **p<0.01, ***p<0.001. In A-D individual experimental replicates are represented. Figure 4—source data 1. Quantification of GFP-positive neurons, RT-qPCR and nuclear staining intensity for .

    Journal: eLife

    Article Title: Neuronal hyperexcitability is a DLK-dependent trigger of herpes simplex virus reactivation that can be induced by IL-1

    doi: 10.7554/eLife.58037

    Figure Lengend Snippet: ( A ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated sodium channel blocker tetrodotoxin (TTX; 1 µM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( B ) Latently infected cultures were reactivated with forskolin in the presence of the voltage-gated potassium channel blocker tetraethylammonium (TEA; 10 mM) and the number of Us11-GFP-positive neurons quantified at 3 days post-reactivation. ( C ) Forskolin-mediated reactivation in the presence of the HCN channel blockers ZD 7288 (10μM) quantified as the numbers of Us11-GFP-positive neurons at 3 days post-reactivation. ( D ) The effect of ZD 7288 on the HSV lytic gene transcript ICP27 during Phase I reactivation measured at 20 hr post-forskolin treatment by RT-qPCR. In A-D individual experimental replicates are represented along with the mean and SEM. ( E and F ) Quantification of the relative nuclear staining for H3K9me3/S10p and γH2AX in SCG neurons at 5 hr post-forskolin treatment and in the presence of ZD 7288 from >800 cells/condition from two independent experiments. Data are plotted around the mean, with the boxes representing the 25 th -75 th percentiles and the whiskers the 5 st -95 th percentiles. Statistical comparisons were made using a one-way ANOVA with a Tukey’s multiple comparison ( A–D ) or two-tailed unpaired t-test ( E–F ). *p<0.05, **p<0.01, ***p<0.001. In A-D individual experimental replicates are represented. Figure 4—source data 1. Quantification of GFP-positive neurons, RT-qPCR and nuclear staining intensity for .

    Article Snippet: Compounds used in the study are as follows: Acycloguanosine, FUDR, Uridine, SP600125, GNE-3511, GSK-J4, L-glutamic acid, and Ivabradine (Millipore Sigma); Forskolin, LY 294002, 666–15, SQ 22536, KT 5720, tetraethylammonium chloride, cesium chloride, OG-L002, S2101, tetrotdotoxin, and ESI-09 (Tocris); 1,9-dideoxy-Forskolin, ZD 7288 and 8-bromo-cyclic AMP (Cayman Chemicals); nerve growth factor 2.5S (Alomone Labs); Primocin (Invivogen); aphidicolin (AG Scientific); IL-1β (Shenandoah Biotechnology); WAY-150138 was kindly provided by Pfizer, Dr. Jay Brown at the University of Virginia, and Dr. Lynn Enquist at Princeton University.

    Techniques: Infection, Quantitative RT-PCR, Staining, Two Tailed Test

    Journal: eLife

    Article Title: Neuronal hyperexcitability is a DLK-dependent trigger of herpes simplex virus reactivation that can be induced by IL-1

    doi: 10.7554/eLife.58037

    Figure Lengend Snippet:

    Article Snippet: Compounds used in the study are as follows: Acycloguanosine, FUDR, Uridine, SP600125, GNE-3511, GSK-J4, L-glutamic acid, and Ivabradine (Millipore Sigma); Forskolin, LY 294002, 666–15, SQ 22536, KT 5720, tetraethylammonium chloride, cesium chloride, OG-L002, S2101, tetrotdotoxin, and ESI-09 (Tocris); 1,9-dideoxy-Forskolin, ZD 7288 and 8-bromo-cyclic AMP (Cayman Chemicals); nerve growth factor 2.5S (Alomone Labs); Primocin (Invivogen); aphidicolin (AG Scientific); IL-1β (Shenandoah Biotechnology); WAY-150138 was kindly provided by Pfizer, Dr. Jay Brown at the University of Virginia, and Dr. Lynn Enquist at Princeton University.

    Techniques: Recombinant, Plasmid Preparation, Blocking Assay, Sequencing, shRNA, SYBR Green Assay, Staining