trek1 antibody  (Alomone Labs)

Journal: bioRxiv

Article Title: Hyperexcitability in Alzheimer’s Disease triggers a compensatory neuroprotective response via TREK1

doi: 10.1101/2025.10.16.682816

Figure Lengend Snippet: a, Representative immunofluorescence images showing elevated TREK1 expression in the hippocampus and cortex of APP/PS1 mice compared to age- matched wild-type controls. b, Quantification of TREK1 fluorescence intensity in the APP/PS1 mice hippocampus (n=9-12 sections; *P < 0.05; two-tailed unpaired t-test). c, Quantification of TREK1 fluorescence intensity in the APP/PS1 mice cortex (n=8 sections; *P < 0.05; two-tailed unpaired t-test). d, Western blot of brain cortex lysates showing increased TREK1 protein abundance in APP/PS1 mice. e, Densitometric analysis and quantification of TREK1 protein levels normalized to GAPDH (n=3; **P < 0.01; two-tailed unpaired t-test). f, Representative immunofluorescence images showing increased TREK1 expression in the hippocampus and cortex of 3xTg mice compared to wild-type controls. g, Quantification of TREK1 fluorescence intensity in the 3xTg mice hippocampus (n=10-14 sections; ****P < 0.0001; two-tailed unpaired t- test). h, Quantification of TREK1 fluorescence intensity in the 3xTg mice cortex (n=8- 9 sections; *P < 0.05; two-tailed unpaired t-test). i, Western blot analysis shows increased TREK1 protein abundance in 3xTg mouse brain homogenates. j, Densitometric analysis and quantification of TREK1 protein levels normalized to GAPDH in 3xTg mice (n=3; ****P < 0.0001; two-tailed unpaired t-test). k, Schematic representation showing increased TREK1 expression in AD transgenic mice. Data are presented as mean ± SEM. 3–4 mice per group were used.

Article Snippet: Primary antibodies- chicken MAP2 (1:1000, Invitrogen, PA1-10005), rabbit TREK1 (1:100, Alomone Labs, #APC-047), Rabbit CTCF (1:100, Invitrogen, #MA5-88115), Mouse AC1 (1:50, Santa Cruz, #SC- 365350), Mouse AC8 (1:50, Santa Cruz, #SC-377442) and rabbit MAP2 (1:150, Cell Signalling Technology, #8707S) were diluted in PBST and applied overnight at 4°C.

Techniques: Immunofluorescence, Expressing, Fluorescence, Two Tailed Test, Western Blot, Quantitative Proteomics, Transgenic Assay

Journal: bioRxiv

Article Title: Hyperexcitability in Alzheimer’s Disease triggers a compensatory neuroprotective response via TREK1

doi: 10.1101/2025.10.16.682816

Figure Lengend Snippet: a, Representative immunofluorescence images showing increased TREK1 expression in primary neurons treated with Aβ42o compared with Aβ42 monomers, Aβ42o, and control. b, Quantification of TREK1 fluorescence intensity upon treatment with Aβ42o/ Aβ42 monomer compared with control (n=12-17 cells; *p < 0.05, one-way ANOVA followed by Šidák’s multiple comparisons test). c, Quantification of TREK1 fluorescence intensity upon treatment with Aβ42o/ Aβ40o compared with control (n=64-103 cells; ****p < 0.0001, ###p < 0.001, one-way ANOVA followed by Šidák’s multiple comparisons test). d, Western blot analysis of primary neuron–astrocyte co-cultures confirming increased TREK1 levels in Aβ42o treated neurons compared with controls. e, Densitometric analysis and quantification of TREK1 protein levels normalized to GAPDH in Aβ42o treated co-cultures (n=3; **p < 0.01, unpaired t-test). f, Hippocampal sections from C57BL/6 wild-type mice 24 hours after intrahippocampal injection of Aβ42o showing increased TREK1 expression compared with vehicle-injected controls. g, Quantification of TREK1 expression from (f) (n=21-27 sections; ****p < 0.0001, unpaired t-test). h, Representative images showing that co-treatment of Aβ42o with APV or TTX reduces Aβ42o-induced TREK1 upregulation in primary cultures. i, Quantification of TREK1 fluorescence intensity in APV/TTX treated primary cultures (n=71-154 cells; ****p < 0.0001, ####p < 0.0001, ††††p < 0.0001, one-way ANOVA with Šidák’s test). j, Exposure to glutamate mimics Aβ42o-induced TREK1 upregulation. k, Quantification of TREK1 fluorescence intensity following glutamate treatment (n=36- 39 cells; **** p < 0.0001, unpaired t-test). l, Treatment with NMDA or KCl to induce neuronal hyperexcitability increases TREK1 levels. m, Quantification of TREK1 fluorescence intensity following KCl and NMDA treatment (n=25-79 cells; ****p < 0.0001, ####p < 0.0001, one-way ANOVA with Šidák’s test). n, Representative immunofluorescence images of brain sections from Aβ42o injected mice showing elevated TREK1 expression in the hippocampus, which is attenuated by co-injection with APV. o, Quantification of hippocampal TREK1 intensity after injecting Aβ42o and/or APV (n=18-27 sections; ****p < 0.0001, one-way ANOVA followed by Šidák’s multiple comparisons test). p, Representative images of hippocampal sections from 3xTg mice treated with APV or vehicle showing decreased TREK1 expression following NMDA receptor blockade. q, Quantification of TREK1 intensity following APV treatment in 3xTg mice (n=10-18 sections; **p < 0.01, unpaired t-test). r, Schematic representation illustrating that Aβ42o induced neuronal hyperexcitability drives increased TREK1 expression. Data are presented as mean ± SEM. 3-5 independent cultures or animals per group were used.

Article Snippet: Primary antibodies- chicken MAP2 (1:1000, Invitrogen, PA1-10005), rabbit TREK1 (1:100, Alomone Labs, #APC-047), Rabbit CTCF (1:100, Invitrogen, #MA5-88115), Mouse AC1 (1:50, Santa Cruz, #SC- 365350), Mouse AC8 (1:50, Santa Cruz, #SC-377442) and rabbit MAP2 (1:150, Cell Signalling Technology, #8707S) were diluted in PBST and applied overnight at 4°C.

Techniques: Immunofluorescence, Expressing, Control, Fluorescence, Western Blot, Injection

Journal: bioRxiv

Article Title: Hyperexcitability in Alzheimer’s Disease triggers a compensatory neuroprotective response via TREK1

doi: 10.1101/2025.10.16.682816

Figure Lengend Snippet: a, Representative immunofluorescence images showing increased TREK1 expression upon Aβo treatment, which is markedly reduced by co-treatment with the calcium chelator BAPTA-AM. b, Quantification of TREK1 fluorescence intensity following Aβ42o and/or BAPTA-AM treatment (n=61-90 Cells; **p < 0.01, ####p < 0.0001; one-way ANOVA with Šidák’s test). c, Representative images showing that TREK1 increase induced by Aβ42o is attenuated by the AC1 inhibitor ST034307. d, Quantification of TREK1 fluorescence intensity following Aβ42o and/or ST034307 treatment (n=23-31 cells; **p < 0.01, ###p < 0.001; one-way ANOVA with Šidák’s test). e, Representative images showing TREK1 expression is decreased in Aβ42o treated neurons transfected with AC1-specific siRNA but not with scrambled (Sc) siRNA. f, Quantification of TREK1 fluorescence intensity following Aβ42o treatment with AC1 knockdown (n=71-128 cells; ****p < 0.0001, ####p < 0.0001; one-way ANOVA with Šidák’s test). g, Representative images showing TREK1 expression is reduced in Aβ42o-treated neurons transfected with AC8-specific siRNA, but not with scrambled siRNA. h, Quantification of TREK1 fluorescence intensity following Aβ42o treatment with AC8 knockdown (n=14-19 cells; ****p < 0.0001, unpaired t-test). i, Treatment with the cAMP analog 8-CPT-cAMP increases TREK1 expression in primary neurons. j, Quantification of TREK1 fluorescence intensity following 8-CPT- cAMP treatment (n=57-62 cells; ***p < 0.001, unpaired t-test). k, Treatment with forskolin, a cAMP activator, mimics Aβ42o by increasing TREK1 expression in primary neurons. l, Quantification of TREK1 fluorescence intensity following forskolin treatment (n=146-242 cells; ****p < 0.0001, ####p < 0.0001; one-way ANOVA with Šidák’s test). m, Schematic representation illustrating that Aβ42o-induced TREK1 upregulation is mediated by calcium influx via the AC1/AC8–cAMP signaling pathway. Data are presented as mean ± SEM. 3-5 independent cultures per group were used.

Article Snippet: Primary antibodies- chicken MAP2 (1:1000, Invitrogen, PA1-10005), rabbit TREK1 (1:100, Alomone Labs, #APC-047), Rabbit CTCF (1:100, Invitrogen, #MA5-88115), Mouse AC1 (1:50, Santa Cruz, #SC- 365350), Mouse AC8 (1:50, Santa Cruz, #SC-377442) and rabbit MAP2 (1:150, Cell Signalling Technology, #8707S) were diluted in PBST and applied overnight at 4°C.

Techniques: Immunofluorescence, Expressing, Fluorescence, Transfection, Knockdown

Journal: bioRxiv

Article Title: Hyperexcitability in Alzheimer’s Disease triggers a compensatory neuroprotective response via TREK1

doi: 10.1101/2025.10.16.682816

Figure Lengend Snippet: a, Representative immunofluorescence images showing increased TREK1 expression in Aβ42o-treated neurons, which is decreased upon co-treatment with the PKA inhibitor KT5720. b, Quantification of TREK1 fluorescence intensity following Aβ42o and/or KT5720 treatment (n = 52-70 cells; ****p < 0.0001, ####p < 0.0001; one-way ANOVA with Šidák’s test). c, Representative images showing that TREK1 increase induced by Aβ42o is attenuated by another PKA inhibitor H89. d, Quantification of TREK1 fluorescence intensity following Aβ42o and/or H89 treatment (n = 43–49 cells; **** p < 0.0001, ####p < 0.0001; one-way ANOVA with Šidák’s test). e, Schematic representation of the TREK1 locus on chromosome 1q41 showing predicted promoters (blue boxes; P1–P3) and ENCODE-annotated CTCF binding sites (red boxes; accession IDs indicated). Genomic positions are indicated relative to the transcription start site (TSS). ReMap ChIP-seq and density tracks demonstrate experimental support for the predicted binding sites, with peaks at site E1421793 located proximal to the promoter (P1). Motif analysis using JASPAR confirmed the presence of a consensus CTCF motif within this region (right panel), with associated FIMO statistics (score, p-value, and q-value). The identified site (highlighted in red) spans chr1:215076354–215076685 (band 1q41), has a genomic size of 332 bp, and is classified as “CTCF-bound” in ENCODE. f, Representative images showing decreased TREK1 expression in Aβ42o treated neurons transfected with CTCF-specific siRNA compared to scrambled (Sc) siRNA. g, Quantification of TREK1 fluorescence intensity following Aβ42o and/or CTCF knockdown (n = 37–76 cells; **** p < 0.0001, #### p < 0.0001; one-way ANOVA with Šidák’s test). h, Representative images showing decreased TREK1 expression in hippocampal neurons of 3xTg mice following intrahippocampal injection of CTCF shRNA lentivirus compared to control. i, Quantification of TREK1 fluorescence intensity in 3xTg mice after CTCF knockdown (n = 13–19 sections; ****p < 0.0001; unpaired t-test). j, Schematic representation of the PKA–CTCF signaling axis in regulating Aβ42- induced TREK1 expression. Data are presented as mean ± SEM. 3-5 independent cultures or animals per group were used.

Article Snippet: Primary antibodies- chicken MAP2 (1:1000, Invitrogen, PA1-10005), rabbit TREK1 (1:100, Alomone Labs, #APC-047), Rabbit CTCF (1:100, Invitrogen, #MA5-88115), Mouse AC1 (1:50, Santa Cruz, #SC- 365350), Mouse AC8 (1:50, Santa Cruz, #SC-377442) and rabbit MAP2 (1:150, Cell Signalling Technology, #8707S) were diluted in PBST and applied overnight at 4°C.

Techniques: Immunofluorescence, Expressing, Fluorescence, Binding Assay, ChIP-sequencing, Transfection, Knockdown, Injection, shRNA, Control

Journal: bioRxiv

Article Title: Hyperexcitability in Alzheimer’s Disease triggers a compensatory neuroprotective response via TREK1

doi: 10.1101/2025.10.16.682816

Figure Lengend Snippet: a, Representative calcium imaging traces from control and Aβ42o treated neurons showing that Aβ42o increases spontaneous calcium transient frequency. This hyperexcitability is further enhanced by the TREK1 inhibitor spadin and suppressed by the TREK1 activator BL-1249. b, Quantification of calcium event frequency upon treatment with Aβ42o/spadin/BL-1249 (n = 512–1002 cells; ****p < 0.0001, ##p < 0.01, †p < 0.05; one-way ANOVA with Šidák’s test). c, Representative FluoVolt traces measuring membrane potential fluctuations manifest enhanced neuronal activity in the presence of TREK1 inhibitor spadin and suppressed neuronal activity by the TREK1 activator BL-1249 compared to Aβ42o treatment alone. d, Quantification of potential spike frequency upon treatment with Aβ42o/spadin/BL- 1249 (n = 31–105 cells; **p < 0.01, #p < 0.05, ††p < 0.01; one-way ANOVA with Šidák’s test). e, Representative calcium traces from neurons treated with Aβ42o along with scrambled (Sc) siRNA, KCNK2 siRNA, or a KCNK2 overexpression (OE) construct. f, Quantification of calcium event frequency upon knocking down KCNK2 in the presence of Aβ42o treatment (n = 49–67 cells; *p < 0.05; unpaired t-test). g, Quantification of calcium event frequency upon overexpressing KCNK2 in the presence of Aβ42o treatment (n = 37–48 cells; *p < 0.05, #p < 0.05; one-way ANOVA with Šidák’s test). h, Representative patch-clamp recordings of action potentials in control, Aβ42o, and Aβ42o + spadin treated neurons showing exacerbated action potential firing with TREK1 blockade. i, Quantification of action potential frequency following Aβ42o and/or spadin treatment (n = 17 cells; *p < 0.05, #p < 0.05; one-way ANOVA with Šidák’s test). j, Resting membrane potential (RMP) is more depolarized in Aβ42o treated neurons compared to control, and further depolarizes in presence of spadin with Aβ42o (n = 19 cells; ****p < 0.0001, #p < 0.05; one-way ANOVA with Šidák’s test). k, Representative traces showing excitatory postsynaptic current (EPSC) frequency is increased in neurons treated with Aβ42o + spadin compared to Aβ42o alone. l, Quantification of EPSC frequency (n = 19 cells; *p < 0.05, ##p < 0.01; one-way ANOVA with Šidák’s test). m, Quantification of EPSC amplitude (n = 19 cells). n, Representative traces showing Inhibitory postsynaptic current (IPSC) frequency is decreased in neurons treated with Aβ42o+spadin compared to Aβ42o alone. o, Quantification of IPSC frequency (n = 10 cells; **p < 0.01, #p < 0.05; one-way ANOVA with Šidák’s test). p, Quantification of IPSC amplitude (n = 10 cells). q, Representative ex vivo calcium imaging heat map from hippocampal slices of 3xTg mice injected with TREK1 shRNA lentivirus showing elevated calcium activity compared to sc shRNA-injected mice. r, Representative calcium imaging traces demonstrating increased calcium transient frequency following TREK1 knockdown. s, Quantification of calcium event frequency in TREK1 knockdown mice compared to sc shRNA-injected mice (n = 13–27 cells; *p < 0.05; unpaired t-test). Data are presented as mean ± SEM from 3-5 independent cultures.

Article Snippet: Primary antibodies- chicken MAP2 (1:1000, Invitrogen, PA1-10005), rabbit TREK1 (1:100, Alomone Labs, #APC-047), Rabbit CTCF (1:100, Invitrogen, #MA5-88115), Mouse AC1 (1:50, Santa Cruz, #SC- 365350), Mouse AC8 (1:50, Santa Cruz, #SC-377442) and rabbit MAP2 (1:150, Cell Signalling Technology, #8707S) were diluted in PBST and applied overnight at 4°C.

Techniques: Imaging, Control, Membrane, Activity Assay, Over Expression, Construct, Patch Clamp, Ex Vivo, Injection, shRNA, Knockdown

Journal: bioRxiv

Article Title: Hyperexcitability in Alzheimer’s Disease triggers a compensatory neuroprotective response via TREK1

doi: 10.1101/2025.10.16.682816

Figure Lengend Snippet: a, Representative immunofluorescence images showing increased VGLUT1 intensity in the hippocampus of 3xTg mice 15 days after intrahippocampal injection with TREK1 shRNA lentivirus compared with vehicle-injected controls. b, Representative images showing decreased VGAT intensity under the same TREK1 knockdown conditions. c, Quantification of VGLUT1 fluorescence intensity in 3xTg mice after TREK1 knockdown (n = 67–76 sections; ****p < 0.0001; unpaired t-test). d, Quantification of VGAT fluorescence intensity in 3xTg mice after TREK1 knockdown (n = 60–63 sections; ****p < 0.0001; unpaired t-test). e, Quantification of Excitatory/inhibitory (E/I) ratio, calculated as VGLUT1/VGAT intensity, is markedly elevated in TREK1 knockdown mice compared with vehicle injected controls (n = 3– 4; **p < 0.01; unpaired t-test). f, Representative immunofluorescence images demonstrating enhanced Aβ deposition in the hippocampus of TREK1 knockdown 3xTg mice compared with vehicle-injected controls. g, Quantification of Aβ fluorescence intensity in 3xTg mice after TREK1 knockdown (n = 50 sections; *p < 0.05; unpaired t-test). h, Quantification showing a significant decrease in MAP2 intensity in the hippocampus of 3xTg mice injected with TREK1 shRNA compared with scrambled (Sc) shRNA controls (n = 159–166 sections; ****p < 0.0001; unpaired t-test). i, Schematic representation illustrating the effects of TREK1 knockdown on excitatory/inhibitory balance. Data are expressed as mean ± SEM from 3–4 mice per group.

Article Snippet: Primary antibodies- chicken MAP2 (1:1000, Invitrogen, PA1-10005), rabbit TREK1 (1:100, Alomone Labs, #APC-047), Rabbit CTCF (1:100, Invitrogen, #MA5-88115), Mouse AC1 (1:50, Santa Cruz, #SC- 365350), Mouse AC8 (1:50, Santa Cruz, #SC-377442) and rabbit MAP2 (1:150, Cell Signalling Technology, #8707S) were diluted in PBST and applied overnight at 4°C.

Techniques: Immunofluorescence, Injection, shRNA, Knockdown, Fluorescence

Journal: bioRxiv

Article Title: Hyperexcitability in Alzheimer’s Disease triggers a compensatory neuroprotective response via TREK1

doi: 10.1101/2025.10.16.682816

Figure Lengend Snippet: The diagram summarizes the signaling cascade by which Aβ42 induced neuronal hyperexcitability drives TREK1 upregulation via calcium influx and the AC1/AC8–cAMP–PKA–CTCF axis. TREK1 upregulation decreases neuronal excitability, limits excitatory/inhibitory balance, thereby improves neuronal health in 3xTg mice.

Article Snippet: Primary antibodies- chicken MAP2 (1:1000, Invitrogen, PA1-10005), rabbit TREK1 (1:100, Alomone Labs, #APC-047), Rabbit CTCF (1:100, Invitrogen, #MA5-88115), Mouse AC1 (1:50, Santa Cruz, #SC- 365350), Mouse AC8 (1:50, Santa Cruz, #SC-377442) and rabbit MAP2 (1:150, Cell Signalling Technology, #8707S) were diluted in PBST and applied overnight at 4°C.

Techniques:

Journal: Nature Communications

Article Title: Pt nanoshells with a high NIR-II photothermal conversion efficiency mediates multimodal neuromodulation against ventricular arrhythmias

doi: 10.1038/s41467-024-50557-w

Figure Lengend Snippet: a Flowchart of calcium imaging assay performed on HT-22 cells. b calcium imaging of HT-22 cells under different experimental conditions, n = 6 biologically independent replicates. c Western blotting for TRPV1 and TREK1 from HT-22 and HEK-293T cells, n = 4 biologically independent replicates. Percentage of d TRPV1 and f TREK1 groups of HT-22 cells within the field of view of the fluorescence microscope that responded to laser stimulation, n = 6 biologically independent replicates. Temporal dynamics of Ca 2+ signals in e TRPV1 and g TREK1 groups of cells. The solid lines indicate the mean, and the shade represents the standard error of the mean (SEM). h Cell viability of HT-22 treated with different concentrations of PtNP-shell for 24 h. Effect of NIR-II laser irradiation with varying durations on the viability of HT-22 cells treated with PtNP-shell (50 μg mL −1 ) (Power densities: i 0.75 W cm −2 and j 1 W cm −2 ), n = 6 biologically independent replicates. The error bar indicates S.E.M. Two-way ANOVA with Tukey’s honestly significant difference (HSD) test was applied for statistical analysis of ( d ) and ( f ). One-way ANOVA with Dunnett’s multiple comparisons test was applied for statistical analysis of ( i ) and ( j ). Source data are provided as a Source Data file.

Article Snippet: Mouse monoclonal anti-TREK1 antibody (Cat. No. sc-398449, Clone No. F-6) used in western blot and immunofluorescence staining was purchased from Santa Cruz Biotechnology (TX, USA).

Techniques: Imaging, Western Blot, Fluorescence, Microscopy, Irradiation

Journal: Nature Communications

Article Title: Pt nanoshells with a high NIR-II photothermal conversion efficiency mediates multimodal neuromodulation against ventricular arrhythmias

doi: 10.1038/s41467-024-50557-w

Figure Lengend Snippet: a Location of the canine LSG. b Schematic illustration of the process of photothermal modulation of LSG. c Temperature curves of LSG under NIR-II laser irradiation. d Typical thermal imaging diagram of photothermally modulated activation of LSG. e Representative images of BP elevation induced after stimulation of LSG with different voltages. Maximal SBP changes of beagle treatment with PtNP-shell or control f before and g after NIR-II exposure, n = 6 biologically independent replicates. h Quantification of the LSG neural activity recordings, n = 6 biologically independent replicates. i Representative immunofluorescent images of TH, c-Fos, and TREK1 in the LSG of beagles following different treatments. Data are shown as the mean ± S.E.M. Unpaired two-tailed Student’s t -test was applied for statistical analysis of ( g ). Two-way ANOVA with Tukey’s HSD test was applied for statistical analysis of ( h ). LSG left stellate ganglion, BP blood pressure, SBP systolic BP. Source data are provided as a Source Data file.

Article Snippet: Mouse monoclonal anti-TREK1 antibody (Cat. No. sc-398449, Clone No. F-6) used in western blot and immunofluorescence staining was purchased from Santa Cruz Biotechnology (TX, USA).

Techniques: Irradiation, Imaging, Activation Assay, Control, Activity Assay, Two Tailed Test