Journal: eLife

Article Title: Natural antisense transcripts regulate the neuronal stress response and excitability

doi: 10.7554/eLife.01849

Figure Lengend Snippet: ( A ) Blocking SEI channel activity in ppk29 mutants with the hERG channel blocker Cisapride eliminate the protective effect in a dose dependent manner (n = 8 per genotype, p<0.01, two-way ANOVA; genotype, dose, and genotype by dose showed significant effects, p =<0.001). ( B ) Schematic representation of transgenic constructs. ( C ) Neuronal expression of ppk29 -3′UTR is sufficient to rescue the majority of the protective effect of the ppk29 mutation (n = 12, p<0.01, one-way ANOVA). Data are presented as mean ± SEM. Different letters above bars represent significantly different groups (Tukey post hoc analysis, p<0.05). ( D ) Neuronal expression of sei cDNA with or without its endogenous 3′UTR, but not the 3′UTR alone , is sufficient to rescue the sei mutation (n = 12, p<0.001, one-way ANOVA). DOI: http://dx.doi.org/10.7554/eLife.01849.015

Article Snippet: Stock solutions of hERG blockers were kept as 10 mM Cisapride (Sigma-Aldrich, St. Louis, MO, USA) in DMSO and 100 mM E 4301 (Alomone labs, Jerusalem, Israel) in distilled water.

Techniques: Blocking Assay, Activity Assay, Transgenic Assay, Construct, Expressing, Mutagenesis

Journal: eLife

Article Title: Natural antisense transcripts regulate the neuronal stress response and excitability

doi: 10.7554/eLife.01849

Figure Lengend Snippet: Treating ppk29 mutants flies with hERG inhibitors cisapride ( A ) and E−4301 ( B ) lead to a significantly faster heat-induced seizures and paralyses in all tested genotypes (n = 8 for each genotype, two-way ANOVA with a Tukey's post-hoc test; the interaction between genotype and concentration is significant for both drugs, p =<0.001). Average data are presented as mean ± SEM. Different letters above bars represent significantly different groups. DOI: http://dx.doi.org/10.7554/eLife.01849.016

Article Snippet: Stock solutions of hERG blockers were kept as 10 mM Cisapride (Sigma-Aldrich, St. Louis, MO, USA) in DMSO and 100 mM E 4301 (Alomone labs, Jerusalem, Israel) in distilled water.

Techniques: Concentration Assay

Journal: EMBO Molecular Medicine

Article Title: A new hERG allosteric modulator rescues genetic and drug‐induced long‐ QT syndrome phenotypes in cardiomyocytes from isogenic pairs of patient induced pluripotent stem cells

doi: 10.15252/emmm.201606260

Figure Lengend Snippet: Expression analysis as measured by RT–qPCR of cardiac ion channel genes and sarcomeric protein TNNT2 gene in undifferentiated hiPSCs and hiPSC‐CMs. Data are expressed as fold change versus. RPL37A . N = 3. * P < 0.05. The colour of the asterisks indicates comparisons and relative statistical significance. QT intervals (left), RR intervals (right) and QTcB intervals measured with MEA in CMs derived from all the control hPSC lines under baseline conditions. N = 17. * P < 0.05. The colour of the asterisks indicates comparisons and relative statistical significance. Data Information: (A) Kruskal–Wallis test with Dunn's multiple comparisons test: hERG 1a/b: JLNS R594Q versus Undiff.: 0.0296; LQT2 corr versus Undiff.: 0.0011. hERG 1a: JLNS R594Q versus Undiff.: 0.0088; LQT2 corr versus Undiff.: 0.0012. KCNE2: LQT2 corr versus Undiff.: 0.0012; LQT2 N996I versus Undiff.: 0.0468. TNNT2: WT versus Undiff.: 0.0458; LQT1 R594Q versus Undiff.: 0.0320; JLNS R594Q versus Undiff.: 0.0052; LQT2 corr versus Undiff.: 0.0444. LQT2 N996I versus Undiff.: 0.0468. LQT1 corr versus Undiff.: 0.0444. LQT1 R190Q versus Undiff.: 0.0493. CACNA1c: LQT1 R594Q versus Undiff.: 0.0076; JLNS R594Q versus Undiff.: 0.0429; LQT2 corr versus Undiff.: 0.0164. KCNJ12: LQT2 corr versus Undiff.: 0.0024; LQT2 corr versus LQT1 corr : 0.0060. (B) One‐way ANOVA with Holm–Sidak's multiple comparison test: QT intervals: WT versus LQT1 corr : < 0.0001; LQT2 corr versus LQT1 corr : 0.0006; LQT1 corr versus hESC WT : < 0.0001. RR intervals: WT versus LQT1 corr : < 0.0001; WT versus LQT2 corr : 0.0009; WT versus hESC WT : < 0.0001. QTc B : WT versus LQT1 corr : 0.0475; LQT1 corr versus LQT2 corr : 0.0432; LQT1 corr versus hESC WT : < 0.0001. LQT2 corr versus hESC WT : 0.0432. (A, B) Data are expressed and plotted as the mean ± SEM. Source data are available online for this figure.

Article Snippet: Asymptomatic or borderline LQTS mutation carriers are highly represented in the LQTS population [36 and 19%, respectively, for LQT1 and LQT2 (Schwartz et al , )], and they are particularly susceptible to hERG blockers (Fitzgerald & Ackerman, ).

Techniques: Expressing, Quantitative RT-PCR, Derivative Assay

Journal: PLoS ONE

Article Title: Toward in vivo -relevant hERG safety assessment and mitigation strategies based on relationships between non-equilibrium blocker binding, three-dimensional channel-blocker interactions, dynamic occupancy, dynamic exposure, and cellular arrhythmia

doi: 10.1371/journal.pone.0234946

Figure Lengend Snippet: Reference compounds studied in this work, including hERG blockers and GDN .

Article Snippet: We proceeded to test whether hERG blockers could potentially bind according to a similar drain-plug paradigm using a ligand-based overlay model that we manually docked in the hERG cryo-EM structure (noting that the intra-cellular hERG blocker moiety would necessarily reside within the outer vestibule, rather than the cytoplasm).

Techniques:

Journal: PLoS ONE

Article Title: Toward in vivo -relevant hERG safety assessment and mitigation strategies based on relationships between non-equilibrium blocker binding, three-dimensional channel-blocker interactions, dynamic occupancy, dynamic exposure, and cellular arrhythmia

doi: 10.1371/journal.pone.0234946

Figure Lengend Snippet: (A) Stereo image of the proposed canonical hERG binding mode (exemplified by terfenadine, shown as a molecular surface), in which blockers straddle between the Y and C regions of the pore (where the C region is comprised of the kink between the S6 and proximal C-linker helices). (B) Same as A, but showing the putative protrusion of the tail region of terfenadine from the pore domain (green surface) into the C-linker-enclosed portion of the outer vestibule. (C) Terfenadine was docked manually into clusters of SiteMap site points (white spheres) described in . The butterfly-shaped diphenylmethane tail moiety of terfenadine is complementary in shape to the C-linker helix (noting that butterfly-shaped bisaryl groups are relatively commonplace among hERG blockers ).

Article Snippet: We proceeded to test whether hERG blockers could potentially bind according to a similar drain-plug paradigm using a ligand-based overlay model that we manually docked in the hERG cryo-EM structure (noting that the intra-cellular hERG blocker moiety would necessarily reside within the outer vestibule, rather than the cytoplasm).

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Toward in vivo -relevant hERG safety assessment and mitigation strategies based on relationships between non-equilibrium blocker binding, three-dimensional channel-blocker interactions, dynamic occupancy, dynamic exposure, and cellular arrhythmia

doi: 10.1371/journal.pone.0234946

Figure Lengend Snippet: (A) Stereo view of the blocker overlay. All of the trappable propafenone analogs either terminate prior to this zone, or project a planar group into it (magenta) that coincides with the pore axis. (B) Zoomed-in view of the putative blocker constriction zone, demonstrating the non-planar conformations of non-trappable blockers in this region. (C) The open constriction zone, comprised of Phe656 (yellow side chains) in hERG, with Tyr652 shown for reference (cyan side chains). (D) The closed constriction zone, comprised of Phe468 (yellow side chains), with Tyr464 shown for reference in EAG1 (cyan side chains).

Article Snippet: We proceeded to test whether hERG blockers could potentially bind according to a similar drain-plug paradigm using a ligand-based overlay model that we manually docked in the hERG cryo-EM structure (noting that the intra-cellular hERG blocker moiety would necessarily reside within the outer vestibule, rather than the cytoplasm).

Techniques: