ns6180  (Tocris)

Journal: Cells

Article Title: Reduction of Ca 2+ Entry by a Specific Block of KCa3.1 Channels Optimizes Cytotoxic Activity of NK Cells against T-ALL Jurkat Cells

doi: 10.3390/cells12162065

Figure Lengend Snippet: List of reagents used in this study. NS: not specified. NA: not applicable. Gibco (Billings, MT, USA); Equitech-Bio (Kerrville, TX, USA); Sigma-Aldrich (Saint Louis, MO, USA); Stemcell (CDMX, Mexico, MX); Biolegend (San Diego, CA, USA); Cytiva (Marlborough, MA, USA); Miltenyi Biotec (Auburn, CA, USA); Invitrogen (Waltham MA, USA) and Tocris (Bristol, UK).

Article Snippet: NS6180 , Tocris , 4864 , 10 mM , DMSO.

Techniques: Concentration Assay, Solvent, Sterility, Isolation, Binding Assay

Journal: EMBO Molecular Medicine

Article Title: Synaptic disruption and CREB‐regulated transcription are restored by K + channel blockers in ALS

doi: 10.15252/emmm.202013131

Figure Lengend Snippet: Both K + channel blockers exert a neuroprotective effect in ALS C9orf72 cultures by increasing the neurite length of mutant MN (one‐way ANOVA followed by Dunnett's multiple comparison test). n = 3 independent treatments with each hiPSC line. Scale bar: 50 µm. Representative confocal image showing autophagic targeting of cytotoxic aggresomes in ALS C9orf72 MN upon K + channel blockade. Scale bar: 5 µm. Dashed line represents the cell soma. Apamin and XE991 decrease the size of aberrant aggresomes, confirming enhanced autophagy degradation (one‐way ANOVA followed by Dunnett's multiple comparison test). n = 3 independent treatments with the ALS C9orf72 I line. Scale bar: 5 µm. K + channel blockers increase the levels of autophagy‐specific phospho‐SQSTM1/p62 S403 in ALS C9orf72 MN (one‐way ANOVA followed by Dunnett's multiple comparison test). n = 3 independent treatments with the ALS C9orf72 I line. The load of aggregated SQSTM1/p62 is also reduced by the treatments in ALS C9orf72 cultures (one‐way ANOVA followed by Dunnett's multiple comparison test). n = 3 independent treatments for each cell line. Scale bar: 20 µm. Treatment with the K + channel blockers reduces the number of GGGGCC toxic RNA foci in ALS C9orf72 MN. n = 3 independent treatments with each hiPSC line (one‐way ANOVA followed by Dunnett's multiple comparison test). Scale bar: 5 µm. Dashed line represents the cell soma. Data information: * P < 0.05; ** P < 0.01; and *** P < 0.001. Error bars represent SEM. Arrow indicates the structure displayed at higher magnification. Exact P ‐values are reported in Appendix Table .

Article Snippet: The K + channel blockers XE991, Apamin, Agitoxin, UK78282, Charybdotoxin and NS6180 (all from Tocris) were tested on primary neurons cultured in 96‐well plates and transduced with AAV9‐poly(GA) 175 ‐EGFP.

Techniques: Mutagenesis, Comparison

Journal: EMBO Molecular Medicine

Article Title: Synaptic disruption and CREB‐regulated transcription are restored by K + channel blockers in ALS

doi: 10.15252/emmm.202013131

Figure Lengend Snippet: A, B Volcano plots obtained by DESeq2 analysis showing the effect on the transcriptome of ALS C9orf72 MN exerted by Apamin and XE991 in the ALS C9orf72 II and ALS C9orf72 III lines. C PASTAA ranking of the top transcription factors activated by Apamin and XE991. Significance was set with the algorithm published in Roider et al . D Venn diagram and showing the shared 263 CREB‐controlled genes significantly altered by treatment with K + channel blockers; the bar plot represents the significantly altered genes involved in neuronal activity and autophagy obtained by DESeq2 analysis. E Apamin and XE991 increase the levels of pCREB S133 in all the ALS C9orf72 lines (one‐way ANOVA followed by Dunnett's multiple comparison test). n = 3 independent treatments with each hiPSC line. Scale bar: 10 µm. F ALS C9orf72 MN showed an increased number of Homer1b/c + synaptic contacts after treatment with the K + channel blockers, which in contrast had no significant effect on Healthy MN. n = 3 independent treatments with each hiPSC line (two‐way ANOVA). Scale bar: 5 µm. G Venn diagram showing the number of proteins significantly altered upon Apamin and XE991 treatment in ALS C9orf72 MN. The lines ALS C9orf72 II and ALS C9orf72 III were used as representative of the ALS genotype. H, I Volcano plot displaying the up‐ and down‐regulation of the proteins altered by Apamin and XE991 ( t ‐test). The blue dots indicate proteins involved in synaptic structure and function. J Up‐regulated GO (biological processes) terms in ALS C9orf72 MN treated with either K + channel blockers (Fisher enrichment test). K Synapse‐ and neuronal processes‐related proteins significantly up‐regulated by both treatments. Data information: * P < 0.05 and ** P < 0.01. Error bars represent SEM. Exact P ‐values are reported in Appendix Table .

Article Snippet: The K + channel blockers XE991, Apamin, Agitoxin, UK78282, Charybdotoxin and NS6180 (all from Tocris) were tested on primary neurons cultured in 96‐well plates and transduced with AAV9‐poly(GA) 175 ‐EGFP.

Techniques: Activity Assay, Comparison

Journal: EMBO Molecular Medicine

Article Title: Synaptic disruption and CREB‐regulated transcription are restored by K + channel blockers in ALS

doi: 10.15252/emmm.202013131

Figure Lengend Snippet: Apamin and XE991 increase the neurite length of ALS TBK1 MN (one‐way ANOVA followed by Dunnett's multiple comparison test). n = 3 independent treatments. Scale bar: 50 µm. Both K + channel blockers reduce the accumulation of aggregated SQSTM1 in TBK1‐mutant cells (one‐way ANOVA followed by Dunnett's multiple comparison test). n = 3 independent treatments. Scale bar: 10 µm. K + channel blockade reduces also the size of cytotoxic aggresomes (Kruskal–Wallis test). n = 3 independent treatments. Scale bar: 5 µm. In Apamin‐ and XE991‐treated cultures, the levels of phosphorylated CREB are significantly higher than in vehicle‐treated ones (one‐way ANOVA followed by Dunnett's multiple comparison test). n = 3 independent treatments. Scale bar: 10 µm. Apamin and XE991 rescue the loss of excitatory synapses also in TBK‐mutant MN (Kruskal–Wallis test). n = 3 independent treatments. Scale bar: 5 µm. Data information: * P < 0.05; ** P < 0.01; and *** P < 0.001. Error bars represent SEM. Exact P ‐values are reported in Appendix Table .

Article Snippet: The K + channel blockers XE991, Apamin, Agitoxin, UK78282, Charybdotoxin and NS6180 (all from Tocris) were tested on primary neurons cultured in 96‐well plates and transduced with AAV9‐poly(GA) 175 ‐EGFP.

Techniques: Comparison, Mutagenesis