rabbit polyclonal anti kir4 1  (Alomone Labs)


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    Alomone Labs rabbit polyclonal anti kir4 1
    (A) Experimental design for (B)–(H). (B) Representative images of maximal projection and 3D reconstruction (IMARIS) of mG + astrocytes. (C and D) Surface area and enclosed volume of mG + astrocytes. Graphed as means of each astrocyte in Aldh1l1-CreER T2 :Sox2 fl/fl mice (left, n = 28 control, 43 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (E–H) Automatic tracing of mG + astrocyte processes by the filament tool of IMARIS and quantifications. Graphed as means of each astrocyte (left, n = 19 control, 17 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (I) Left, experimental design for (J)–(P); right, immunofluorescence of SOX2 and GFAP and quantification. n = 4 control, 4 Sox2 icKO. (J) Ctx astrocytes visualized by SR101 (arrows) for whole-cell recordings. (K) Voltage steps for astrocyte recording, from −180 to +20 mV with a step size of 10 mV. (L and M) Representative current tracing (L) and I-V curve (M) of one Sox2 -deficient and one Sox2 -intact astrocyte. (N1–N3) Quantification of cell capacitance (N1, n = 23 control, 18 Sox2 icKO), input resistance (N2, n = 20 control, 19 Sox2 icKO), and resting membrane potential (N3, n = 20 control, 17 Sox2 icKO) of Ctx astrocytes. (O) Western blot and quantification of <t>Kir4.1</t> in the brain. n = 4 control, 4 Sox2 icKO. (P) Representative images and quantification of Kir4.1 intensity in the cortex. n = 4 control, 4 Sox2 icKO. Error bars indicate means ± SEM. Unpaired two-tailed Student’s t test was used for statistically analyzing two groups of data. Please see for statistics. n, biological replicates. Scale bars, (B, E, I, J, and P) 20 μm.
    Rabbit Polyclonal Anti Kir4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti kir4 1/product/Alomone Labs
    Average 96 stars, based on 1 article reviews
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    Images

    1) Product Images from "SOX2 is essential for astrocyte maturation and its deletion leads to hyperactive behavior in mice"

    Article Title: SOX2 is essential for astrocyte maturation and its deletion leads to hyperactive behavior in mice

    Journal: Cell reports

    doi: 10.1016/j.celrep.2022.111842

    (A) Experimental design for (B)–(H). (B) Representative images of maximal projection and 3D reconstruction (IMARIS) of mG + astrocytes. (C and D) Surface area and enclosed volume of mG + astrocytes. Graphed as means of each astrocyte in Aldh1l1-CreER T2 :Sox2 fl/fl mice (left, n = 28 control, 43 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (E–H) Automatic tracing of mG + astrocyte processes by the filament tool of IMARIS and quantifications. Graphed as means of each astrocyte (left, n = 19 control, 17 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (I) Left, experimental design for (J)–(P); right, immunofluorescence of SOX2 and GFAP and quantification. n = 4 control, 4 Sox2 icKO. (J) Ctx astrocytes visualized by SR101 (arrows) for whole-cell recordings. (K) Voltage steps for astrocyte recording, from −180 to +20 mV with a step size of 10 mV. (L and M) Representative current tracing (L) and I-V curve (M) of one Sox2 -deficient and one Sox2 -intact astrocyte. (N1–N3) Quantification of cell capacitance (N1, n = 23 control, 18 Sox2 icKO), input resistance (N2, n = 20 control, 19 Sox2 icKO), and resting membrane potential (N3, n = 20 control, 17 Sox2 icKO) of Ctx astrocytes. (O) Western blot and quantification of Kir4.1 in the brain. n = 4 control, 4 Sox2 icKO. (P) Representative images and quantification of Kir4.1 intensity in the cortex. n = 4 control, 4 Sox2 icKO. Error bars indicate means ± SEM. Unpaired two-tailed Student’s t test was used for statistically analyzing two groups of data. Please see for statistics. n, biological replicates. Scale bars, (B, E, I, J, and P) 20 μm.
    Figure Legend Snippet: (A) Experimental design for (B)–(H). (B) Representative images of maximal projection and 3D reconstruction (IMARIS) of mG + astrocytes. (C and D) Surface area and enclosed volume of mG + astrocytes. Graphed as means of each astrocyte in Aldh1l1-CreER T2 :Sox2 fl/fl mice (left, n = 28 control, 43 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (E–H) Automatic tracing of mG + astrocyte processes by the filament tool of IMARIS and quantifications. Graphed as means of each astrocyte (left, n = 19 control, 17 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (I) Left, experimental design for (J)–(P); right, immunofluorescence of SOX2 and GFAP and quantification. n = 4 control, 4 Sox2 icKO. (J) Ctx astrocytes visualized by SR101 (arrows) for whole-cell recordings. (K) Voltage steps for astrocyte recording, from −180 to +20 mV with a step size of 10 mV. (L and M) Representative current tracing (L) and I-V curve (M) of one Sox2 -deficient and one Sox2 -intact astrocyte. (N1–N3) Quantification of cell capacitance (N1, n = 23 control, 18 Sox2 icKO), input resistance (N2, n = 20 control, 19 Sox2 icKO), and resting membrane potential (N3, n = 20 control, 17 Sox2 icKO) of Ctx astrocytes. (O) Western blot and quantification of Kir4.1 in the brain. n = 4 control, 4 Sox2 icKO. (P) Representative images and quantification of Kir4.1 intensity in the cortex. n = 4 control, 4 Sox2 icKO. Error bars indicate means ± SEM. Unpaired two-tailed Student’s t test was used for statistically analyzing two groups of data. Please see for statistics. n, biological replicates. Scale bars, (B, E, I, J, and P) 20 μm.

    Techniques Used: Immunofluorescence, Western Blot, Two Tailed Test

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Protein Extraction, Recombinant, Electron Microscopy, Injection, Magnetic Cell Separation, Lysis, Bicinchoninic Acid Protein Assay, Western Blot, Glutamate Assay, Labeling, SYBR Green Assay, Chromatin Immunoprecipitation, Software, Activity Assay

    rabbit polyclonal anti kir4 1  (Alomone Labs)


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    Alomone Labs rabbit polyclonal anti kir4 1
    RACK1 KO in astrocytes leads to higher levels of <t>Kir4.1</t> in astrocyte somata and PAPs (A) Generation of a mouse line with RACK1 KO in astrocytes (RACK1 cKO). Shown is a schematic of the RACK1 fl/fl and Aldh1l1-Cre/ERT2 alleles. Deletion of exon 2 in Gnb2l1 (the gene coding for RACK1) is induced in astrocytes by tamoxifen injection; this results in a frameshift and premature termination of Gnb2l1 translation. Primers are indicated by red arrows. (B) PCR assays for Gnb2l1 KO in brain DNA from RACK1 fl/fl or Aldh1l1-CreERT2: RACK1 fl/fl tamoxifen-injected mice (RACK1 cKO). The 898-bp band corresponds to the floxed allele ( <xref ref-type=Table S4 ). The 672-bp band corresponds to the exon 2-deleted allele. (C) Confocal images of RACK1 immunofluorescence (red) in the hippocampus in RACK1 fl/fl and RACK1 cKO mice. Astrocytes are co-immunolabeled for GFAP (green). Some astrocytes are indicated by white arrowheads. Nuclei are stained with DAPI. The bottom panel gives a higher-magnification view of the boxed areas in the RACK1 fl/fl and RACK1 cKO images, which shows that RACK1 is specifically depleted in astrocytes ( ∗ ) and is still expressed by neurons (°). Scale bars, 20 μm. (D), Western blot detection and analysis of Kir4.1 and GLT-1 in protein extracts from whole brain, hippocampus, whole-brain synaptogliosomes, hippocampal synaptogliosomes, or whole-brain microvessels purified from RACK1 fl/fl or RACK1 cKO mice. The position of molecular weight markers is indicated on the right. Signals were normalized against that of stain-free membranes except for the experiment on purified microvessels, where histone 3 was used. The data are quoted as the mean ± SD (N = 5 samples per genotype, 1 mouse per sample); two-tailed unpaired t test or Mann-Whitney test. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 and Figure S2 . " width="250" height="auto" />
    Rabbit Polyclonal Anti Kir4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti kir4 1/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti kir4 1 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "The ribosome-associated protein RACK1 represses Kir4.1 translation in astrocytes and influences neuronal activity"

    Article Title: The ribosome-associated protein RACK1 represses Kir4.1 translation in astrocytes and influences neuronal activity

    Journal: Cell Reports

    doi: 10.1016/j.celrep.2023.112456

    RACK1 KO in astrocytes leads to higher levels of Kir4.1 in astrocyte somata and PAPs (A) Generation of a mouse line with RACK1 KO in astrocytes (RACK1 cKO). Shown is a schematic of the RACK1 fl/fl and Aldh1l1-Cre/ERT2 alleles. Deletion of exon 2 in Gnb2l1 (the gene coding for RACK1) is induced in astrocytes by tamoxifen injection; this results in a frameshift and premature termination of Gnb2l1 translation. Primers are indicated by red arrows. (B) PCR assays for Gnb2l1 KO in brain DNA from RACK1 fl/fl or Aldh1l1-CreERT2: RACK1 fl/fl tamoxifen-injected mice (RACK1 cKO). The 898-bp band corresponds to the floxed allele ( <xref ref-type=Table S4 ). The 672-bp band corresponds to the exon 2-deleted allele. (C) Confocal images of RACK1 immunofluorescence (red) in the hippocampus in RACK1 fl/fl and RACK1 cKO mice. Astrocytes are co-immunolabeled for GFAP (green). Some astrocytes are indicated by white arrowheads. Nuclei are stained with DAPI. The bottom panel gives a higher-magnification view of the boxed areas in the RACK1 fl/fl and RACK1 cKO images, which shows that RACK1 is specifically depleted in astrocytes ( ∗ ) and is still expressed by neurons (°). Scale bars, 20 μm. (D), Western blot detection and analysis of Kir4.1 and GLT-1 in protein extracts from whole brain, hippocampus, whole-brain synaptogliosomes, hippocampal synaptogliosomes, or whole-brain microvessels purified from RACK1 fl/fl or RACK1 cKO mice. The position of molecular weight markers is indicated on the right. Signals were normalized against that of stain-free membranes except for the experiment on purified microvessels, where histone 3 was used. The data are quoted as the mean ± SD (N = 5 samples per genotype, 1 mouse per sample); two-tailed unpaired t test or Mann-Whitney test. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 and Figure S2 . " title="... KO in astrocytes leads to higher levels of Kir4.1 in astrocyte somata and PAPs (A) Generation of ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: RACK1 KO in astrocytes leads to higher levels of Kir4.1 in astrocyte somata and PAPs (A) Generation of a mouse line with RACK1 KO in astrocytes (RACK1 cKO). Shown is a schematic of the RACK1 fl/fl and Aldh1l1-Cre/ERT2 alleles. Deletion of exon 2 in Gnb2l1 (the gene coding for RACK1) is induced in astrocytes by tamoxifen injection; this results in a frameshift and premature termination of Gnb2l1 translation. Primers are indicated by red arrows. (B) PCR assays for Gnb2l1 KO in brain DNA from RACK1 fl/fl or Aldh1l1-CreERT2: RACK1 fl/fl tamoxifen-injected mice (RACK1 cKO). The 898-bp band corresponds to the floxed allele ( Table S4 ). The 672-bp band corresponds to the exon 2-deleted allele. (C) Confocal images of RACK1 immunofluorescence (red) in the hippocampus in RACK1 fl/fl and RACK1 cKO mice. Astrocytes are co-immunolabeled for GFAP (green). Some astrocytes are indicated by white arrowheads. Nuclei are stained with DAPI. The bottom panel gives a higher-magnification view of the boxed areas in the RACK1 fl/fl and RACK1 cKO images, which shows that RACK1 is specifically depleted in astrocytes ( ∗ ) and is still expressed by neurons (°). Scale bars, 20 μm. (D), Western blot detection and analysis of Kir4.1 and GLT-1 in protein extracts from whole brain, hippocampus, whole-brain synaptogliosomes, hippocampal synaptogliosomes, or whole-brain microvessels purified from RACK1 fl/fl or RACK1 cKO mice. The position of molecular weight markers is indicated on the right. Signals were normalized against that of stain-free membranes except for the experiment on purified microvessels, where histone 3 was used. The data are quoted as the mean ± SD (N = 5 samples per genotype, 1 mouse per sample); two-tailed unpaired t test or Mann-Whitney test. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 and Figure S2 .

    Techniques Used: Injection, Immunofluorescence, Immunolabeling, Staining, Western Blot, Purification, Molecular Weight, Two Tailed Test, MANN-WHITNEY

    Absence of RACK1 in astrocytes leads to higher Kir4.1-mediated astrocytic inward K + currents and cell volumes (A) Schematic of electrode positions used to record astrocyte whole-cell currents evoked by Schaffer collateral (SC) stimulation in the CA1 region of hippocampal slices. (B) Left: representative traces of astrocytic whole-cell currents induced by 150-ms voltage steps (from −200 mV to +100 mV, 10-mV steps; black traces at the bottom) in RACK1 fl/fl and RACK1 cKO mice before (black and pink) and after (blue) application of a KIR 4.1 antagonist (VU). Scale bars, 50 ms, 5 nA. Right: current-voltage (I-V) plot in RACK1 fl/fl (white filled dots) and in RACK1 cKO (pink-filled dots) mice before (black) and after (blue) application of VU (N = 7 and 10 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons) The data are quoted as the mean ± SD. (C) Left: representative astrocytic Kir4.1 (VU-sensitive) currents induced by SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (black) and RACK1 cKO (pink) mice. Scale bars, 200 ms, 50 pA. Right: quantification of astrocytic Kir4.1 current peak amplitude after each stimulus during SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (white filled dots) and RACK1 cKO (pink filled dots) mice (N = 6 and 5 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons). The data are quoted as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (D) Illustration of the imaging method with a representative raw confocal image of an isolated RACK1 fl/fl CA1 astrocyte expressing tdTomato. (E–G) Imaris analysis: filament tracing (E), convex hull volume (F), and a 3D Sholl analysis (G). (H) Mean territory volume and filament length of RACK1 fl/fl and RACK1 cKO astrocytes. Shown is a histogram of the data, presented as the mean ± SD (N = 4 mice per genotype, 45 astrocytes); two-tailed t test. The data are quoted as the mean ± SD. (I) A Sholl analysis of the ramification complexity of RACK1 fl/fl and RACK1 cKO astrocytes. Two-way analysis of variance. ∗ p < 0.05, ∗∗ p < 0.01. The data are quoted as the mean ± SD. The raw data are presented in <xref ref-type=Table S3 . " title="Absence of RACK1 in astrocytes leads to higher Kir4.1-mediated astrocytic inward K + currents and cell volumes ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Absence of RACK1 in astrocytes leads to higher Kir4.1-mediated astrocytic inward K + currents and cell volumes (A) Schematic of electrode positions used to record astrocyte whole-cell currents evoked by Schaffer collateral (SC) stimulation in the CA1 region of hippocampal slices. (B) Left: representative traces of astrocytic whole-cell currents induced by 150-ms voltage steps (from −200 mV to +100 mV, 10-mV steps; black traces at the bottom) in RACK1 fl/fl and RACK1 cKO mice before (black and pink) and after (blue) application of a KIR 4.1 antagonist (VU). Scale bars, 50 ms, 5 nA. Right: current-voltage (I-V) plot in RACK1 fl/fl (white filled dots) and in RACK1 cKO (pink-filled dots) mice before (black) and after (blue) application of VU (N = 7 and 10 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons) The data are quoted as the mean ± SD. (C) Left: representative astrocytic Kir4.1 (VU-sensitive) currents induced by SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (black) and RACK1 cKO (pink) mice. Scale bars, 200 ms, 50 pA. Right: quantification of astrocytic Kir4.1 current peak amplitude after each stimulus during SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (white filled dots) and RACK1 cKO (pink filled dots) mice (N = 6 and 5 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons). The data are quoted as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (D) Illustration of the imaging method with a representative raw confocal image of an isolated RACK1 fl/fl CA1 astrocyte expressing tdTomato. (E–G) Imaris analysis: filament tracing (E), convex hull volume (F), and a 3D Sholl analysis (G). (H) Mean territory volume and filament length of RACK1 fl/fl and RACK1 cKO astrocytes. Shown is a histogram of the data, presented as the mean ± SD (N = 4 mice per genotype, 45 astrocytes); two-tailed t test. The data are quoted as the mean ± SD. (I) A Sholl analysis of the ramification complexity of RACK1 fl/fl and RACK1 cKO astrocytes. Two-way analysis of variance. ∗ p < 0.05, ∗∗ p < 0.01. The data are quoted as the mean ± SD. The raw data are presented in Table S3 .

    Techniques Used: Imaging, Isolation, Expressing, Two Tailed Test

    Absence of RACK1 in astrocytes alters network population activity and neuronal responses to intense stimulation (A) Schematic of electrode positions used to record field excitatory postsynaptic potentials (fEPSPs) evoked by SC stimulation in the CA1 region of hippocampal slices. (B) Input-output curves for basal synaptic transmission. Left: representative recordings in RACK1 fl/fl mice (black) and RACK1 cKO mice before (pink) and after (blue) application of a Kir 4.1 antagonist (VU). Scale bars, 10 ms, 0.5 mV. Right: quantification of the fEPSP slope for different fiber volley amplitudes after SC stimulation. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 slices from 4 mice; p = 0.0087; RACK1 cKO: n = 5 slices from 5 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (C) Top: a representative recording of fEPSPs evoked by repetitive stimulation (10 Hz, 30 s) of CA1 SCs in RACK1 fl/fl mice under control conditions. Scale bars, 5 s, 0.2 mV. Bottom: enlarged view of fEPSPs evoked by the first 10 stimuli. Scale bars, 200 ms, 0.2 mV. (D) Changes in the fEPSP slope induced by 10-Hz stimulation relative to responses measured before the onset of stimulation (baseline responses) in RACK1 fl/fl mice (white filled dots) and in RACK1 cKO mice (pink-filled dots) before (black) and after (blue) application of VU. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 from 5 mice; RACK1 cKO: n = 6 slices from 4 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (E) Schematic (left) and picture (right) of a hippocampal slice placed on a multielectrode array (MEA). Scale bar, 200 μm. (F) Representative MEA recordings of burst activity induced in hippocampal slices of RACK1 fl/fl (black) and RACK1 cKO (pink) mice by incubation in Mg 2+ -free ACSF containing 6 mM KCl. The expanded recordings of the bursts (surrounded by gray rectangles) are shown on the right. Scale bars, 10 s (left)/200 ms (right), 50 μV. (G) Quantification of burst frequency (top) and burst duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices. The data are quoted as the mean ± SD. n = 15 slices from 5 mice for RACK1 fl/fl and n = 18 slices from 6 mice for RACK1 cKO; unpaired t test. (H) Representative MEA recordings of hippocampal bursting activity in RACK1 fl/fl (top) and RACK1 cKO (bottom) slices in control (Ct) and during 25-min treatment with the Kir4.1 blocker VU0134992 (VU). The corresponding time-frequency plots are shown under the traces. Scale bar, 20 s/2 min, 50 μV. (I) Quantification of VU’s effect on burst frequency (top) and duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices (RACK1 fl/fl: n = 15 slices from 5 mice for burst frequency and duration, respectively; RACK1 cKO: n = 18 slices from 6 mice for burst frequency and duration; paired t test. The data are quoted as the mean ± SD. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in <xref ref-type=Table S3 . " title="... control (Ct) and during 25-min treatment with the Kir4.1 blocker VU0134992 (VU). The corresponding time-frequency plots are ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Absence of RACK1 in astrocytes alters network population activity and neuronal responses to intense stimulation (A) Schematic of electrode positions used to record field excitatory postsynaptic potentials (fEPSPs) evoked by SC stimulation in the CA1 region of hippocampal slices. (B) Input-output curves for basal synaptic transmission. Left: representative recordings in RACK1 fl/fl mice (black) and RACK1 cKO mice before (pink) and after (blue) application of a Kir 4.1 antagonist (VU). Scale bars, 10 ms, 0.5 mV. Right: quantification of the fEPSP slope for different fiber volley amplitudes after SC stimulation. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 slices from 4 mice; p = 0.0087; RACK1 cKO: n = 5 slices from 5 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (C) Top: a representative recording of fEPSPs evoked by repetitive stimulation (10 Hz, 30 s) of CA1 SCs in RACK1 fl/fl mice under control conditions. Scale bars, 5 s, 0.2 mV. Bottom: enlarged view of fEPSPs evoked by the first 10 stimuli. Scale bars, 200 ms, 0.2 mV. (D) Changes in the fEPSP slope induced by 10-Hz stimulation relative to responses measured before the onset of stimulation (baseline responses) in RACK1 fl/fl mice (white filled dots) and in RACK1 cKO mice (pink-filled dots) before (black) and after (blue) application of VU. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 from 5 mice; RACK1 cKO: n = 6 slices from 4 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (E) Schematic (left) and picture (right) of a hippocampal slice placed on a multielectrode array (MEA). Scale bar, 200 μm. (F) Representative MEA recordings of burst activity induced in hippocampal slices of RACK1 fl/fl (black) and RACK1 cKO (pink) mice by incubation in Mg 2+ -free ACSF containing 6 mM KCl. The expanded recordings of the bursts (surrounded by gray rectangles) are shown on the right. Scale bars, 10 s (left)/200 ms (right), 50 μV. (G) Quantification of burst frequency (top) and burst duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices. The data are quoted as the mean ± SD. n = 15 slices from 5 mice for RACK1 fl/fl and n = 18 slices from 6 mice for RACK1 cKO; unpaired t test. (H) Representative MEA recordings of hippocampal bursting activity in RACK1 fl/fl (top) and RACK1 cKO (bottom) slices in control (Ct) and during 25-min treatment with the Kir4.1 blocker VU0134992 (VU). The corresponding time-frequency plots are shown under the traces. Scale bar, 20 s/2 min, 50 μV. (I) Quantification of VU’s effect on burst frequency (top) and duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices (RACK1 fl/fl: n = 15 slices from 5 mice for burst frequency and duration, respectively; RACK1 cKO: n = 18 slices from 6 mice for burst frequency and duration; paired t test. The data are quoted as the mean ± SD. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 .

    Techniques Used: Activity Assay, Transmission Assay, Incubation


    Figure Legend Snippet:

    Techniques Used: Transduction, Recombinant, Multiplex Assay, Mass Spectrometry, Software

    rabbit polyclonal anti kir4 1 extracellular  (Alomone Labs)


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    Alomone Labs rabbit polyclonal anti kir4 1 extracellular
    RACK1 KO in astrocytes leads to higher levels of <t>Kir4.1</t> in astrocyte somata and PAPs (A) Generation of a mouse line with RACK1 KO in astrocytes (RACK1 cKO). Shown is a schematic of the RACK1 fl/fl and Aldh1l1-Cre/ERT2 alleles. Deletion of exon 2 in Gnb2l1 (the gene coding for RACK1) is induced in astrocytes by tamoxifen injection; this results in a frameshift and premature termination of Gnb2l1 translation. Primers are indicated by red arrows. (B) PCR assays for Gnb2l1 KO in brain DNA from RACK1 fl/fl or Aldh1l1-CreERT2: RACK1 fl/fl tamoxifen-injected mice (RACK1 cKO). The 898-bp band corresponds to the floxed allele ( <xref ref-type=Table S4 ). The 672-bp band corresponds to the exon 2-deleted allele. (C) Confocal images of RACK1 immunofluorescence (red) in the hippocampus in RACK1 fl/fl and RACK1 cKO mice. Astrocytes are co-immunolabeled for GFAP (green). Some astrocytes are indicated by white arrowheads. Nuclei are stained with DAPI. The bottom panel gives a higher-magnification view of the boxed areas in the RACK1 fl/fl and RACK1 cKO images, which shows that RACK1 is specifically depleted in astrocytes ( ∗ ) and is still expressed by neurons (°). Scale bars, 20 μm. (D), Western blot detection and analysis of Kir4.1 and GLT-1 in protein extracts from whole brain, hippocampus, whole-brain synaptogliosomes, hippocampal synaptogliosomes, or whole-brain microvessels purified from RACK1 fl/fl or RACK1 cKO mice. The position of molecular weight markers is indicated on the right. Signals were normalized against that of stain-free membranes except for the experiment on purified microvessels, where histone 3 was used. The data are quoted as the mean ± SD (N = 5 samples per genotype, 1 mouse per sample); two-tailed unpaired t test or Mann-Whitney test. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 and Figure S2 . " width="250" height="auto" />
    Rabbit Polyclonal Anti Kir4 1 Extracellular, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti kir4 1 extracellular/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti kir4 1 extracellular - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "The ribosome-associated protein RACK1 represses Kir4.1 translation in astrocytes and influences neuronal activity"

    Article Title: The ribosome-associated protein RACK1 represses Kir4.1 translation in astrocytes and influences neuronal activity

    Journal: Cell Reports

    doi: 10.1016/j.celrep.2023.112456

    RACK1 KO in astrocytes leads to higher levels of Kir4.1 in astrocyte somata and PAPs (A) Generation of a mouse line with RACK1 KO in astrocytes (RACK1 cKO). Shown is a schematic of the RACK1 fl/fl and Aldh1l1-Cre/ERT2 alleles. Deletion of exon 2 in Gnb2l1 (the gene coding for RACK1) is induced in astrocytes by tamoxifen injection; this results in a frameshift and premature termination of Gnb2l1 translation. Primers are indicated by red arrows. (B) PCR assays for Gnb2l1 KO in brain DNA from RACK1 fl/fl or Aldh1l1-CreERT2: RACK1 fl/fl tamoxifen-injected mice (RACK1 cKO). The 898-bp band corresponds to the floxed allele ( <xref ref-type=Table S4 ). The 672-bp band corresponds to the exon 2-deleted allele. (C) Confocal images of RACK1 immunofluorescence (red) in the hippocampus in RACK1 fl/fl and RACK1 cKO mice. Astrocytes are co-immunolabeled for GFAP (green). Some astrocytes are indicated by white arrowheads. Nuclei are stained with DAPI. The bottom panel gives a higher-magnification view of the boxed areas in the RACK1 fl/fl and RACK1 cKO images, which shows that RACK1 is specifically depleted in astrocytes ( ∗ ) and is still expressed by neurons (°). Scale bars, 20 μm. (D), Western blot detection and analysis of Kir4.1 and GLT-1 in protein extracts from whole brain, hippocampus, whole-brain synaptogliosomes, hippocampal synaptogliosomes, or whole-brain microvessels purified from RACK1 fl/fl or RACK1 cKO mice. The position of molecular weight markers is indicated on the right. Signals were normalized against that of stain-free membranes except for the experiment on purified microvessels, where histone 3 was used. The data are quoted as the mean ± SD (N = 5 samples per genotype, 1 mouse per sample); two-tailed unpaired t test or Mann-Whitney test. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 and Figure S2 . " title="... KO in astrocytes leads to higher levels of Kir4.1 in astrocyte somata and PAPs (A) Generation of ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: RACK1 KO in astrocytes leads to higher levels of Kir4.1 in astrocyte somata and PAPs (A) Generation of a mouse line with RACK1 KO in astrocytes (RACK1 cKO). Shown is a schematic of the RACK1 fl/fl and Aldh1l1-Cre/ERT2 alleles. Deletion of exon 2 in Gnb2l1 (the gene coding for RACK1) is induced in astrocytes by tamoxifen injection; this results in a frameshift and premature termination of Gnb2l1 translation. Primers are indicated by red arrows. (B) PCR assays for Gnb2l1 KO in brain DNA from RACK1 fl/fl or Aldh1l1-CreERT2: RACK1 fl/fl tamoxifen-injected mice (RACK1 cKO). The 898-bp band corresponds to the floxed allele ( Table S4 ). The 672-bp band corresponds to the exon 2-deleted allele. (C) Confocal images of RACK1 immunofluorescence (red) in the hippocampus in RACK1 fl/fl and RACK1 cKO mice. Astrocytes are co-immunolabeled for GFAP (green). Some astrocytes are indicated by white arrowheads. Nuclei are stained with DAPI. The bottom panel gives a higher-magnification view of the boxed areas in the RACK1 fl/fl and RACK1 cKO images, which shows that RACK1 is specifically depleted in astrocytes ( ∗ ) and is still expressed by neurons (°). Scale bars, 20 μm. (D), Western blot detection and analysis of Kir4.1 and GLT-1 in protein extracts from whole brain, hippocampus, whole-brain synaptogliosomes, hippocampal synaptogliosomes, or whole-brain microvessels purified from RACK1 fl/fl or RACK1 cKO mice. The position of molecular weight markers is indicated on the right. Signals were normalized against that of stain-free membranes except for the experiment on purified microvessels, where histone 3 was used. The data are quoted as the mean ± SD (N = 5 samples per genotype, 1 mouse per sample); two-tailed unpaired t test or Mann-Whitney test. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 and Figure S2 .

    Techniques Used: Injection, Immunofluorescence, Immunolabeling, Staining, Western Blot, Purification, Molecular Weight, Two Tailed Test, MANN-WHITNEY

    Absence of RACK1 in astrocytes leads to higher Kir4.1-mediated astrocytic inward K + currents and cell volumes (A) Schematic of electrode positions used to record astrocyte whole-cell currents evoked by Schaffer collateral (SC) stimulation in the CA1 region of hippocampal slices. (B) Left: representative traces of astrocytic whole-cell currents induced by 150-ms voltage steps (from −200 mV to +100 mV, 10-mV steps; black traces at the bottom) in RACK1 fl/fl and RACK1 cKO mice before (black and pink) and after (blue) application of a KIR 4.1 antagonist (VU). Scale bars, 50 ms, 5 nA. Right: current-voltage (I-V) plot in RACK1 fl/fl (white filled dots) and in RACK1 cKO (pink-filled dots) mice before (black) and after (blue) application of VU (N = 7 and 10 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons) The data are quoted as the mean ± SD. (C) Left: representative astrocytic Kir4.1 (VU-sensitive) currents induced by SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (black) and RACK1 cKO (pink) mice. Scale bars, 200 ms, 50 pA. Right: quantification of astrocytic Kir4.1 current peak amplitude after each stimulus during SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (white filled dots) and RACK1 cKO (pink filled dots) mice (N = 6 and 5 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons). The data are quoted as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (D) Illustration of the imaging method with a representative raw confocal image of an isolated RACK1 fl/fl CA1 astrocyte expressing tdTomato. (E–G) Imaris analysis: filament tracing (E), convex hull volume (F), and a 3D Sholl analysis (G). (H) Mean territory volume and filament length of RACK1 fl/fl and RACK1 cKO astrocytes. Shown is a histogram of the data, presented as the mean ± SD (N = 4 mice per genotype, 45 astrocytes); two-tailed t test. The data are quoted as the mean ± SD. (I) A Sholl analysis of the ramification complexity of RACK1 fl/fl and RACK1 cKO astrocytes. Two-way analysis of variance. ∗ p < 0.05, ∗∗ p < 0.01. The data are quoted as the mean ± SD. The raw data are presented in <xref ref-type=Table S3 . " title="Absence of RACK1 in astrocytes leads to higher Kir4.1-mediated astrocytic inward K + currents and cell volumes ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Absence of RACK1 in astrocytes leads to higher Kir4.1-mediated astrocytic inward K + currents and cell volumes (A) Schematic of electrode positions used to record astrocyte whole-cell currents evoked by Schaffer collateral (SC) stimulation in the CA1 region of hippocampal slices. (B) Left: representative traces of astrocytic whole-cell currents induced by 150-ms voltage steps (from −200 mV to +100 mV, 10-mV steps; black traces at the bottom) in RACK1 fl/fl and RACK1 cKO mice before (black and pink) and after (blue) application of a KIR 4.1 antagonist (VU). Scale bars, 50 ms, 5 nA. Right: current-voltage (I-V) plot in RACK1 fl/fl (white filled dots) and in RACK1 cKO (pink-filled dots) mice before (black) and after (blue) application of VU (N = 7 and 10 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons) The data are quoted as the mean ± SD. (C) Left: representative astrocytic Kir4.1 (VU-sensitive) currents induced by SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (black) and RACK1 cKO (pink) mice. Scale bars, 200 ms, 50 pA. Right: quantification of astrocytic Kir4.1 current peak amplitude after each stimulus during SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (white filled dots) and RACK1 cKO (pink filled dots) mice (N = 6 and 5 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons). The data are quoted as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (D) Illustration of the imaging method with a representative raw confocal image of an isolated RACK1 fl/fl CA1 astrocyte expressing tdTomato. (E–G) Imaris analysis: filament tracing (E), convex hull volume (F), and a 3D Sholl analysis (G). (H) Mean territory volume and filament length of RACK1 fl/fl and RACK1 cKO astrocytes. Shown is a histogram of the data, presented as the mean ± SD (N = 4 mice per genotype, 45 astrocytes); two-tailed t test. The data are quoted as the mean ± SD. (I) A Sholl analysis of the ramification complexity of RACK1 fl/fl and RACK1 cKO astrocytes. Two-way analysis of variance. ∗ p < 0.05, ∗∗ p < 0.01. The data are quoted as the mean ± SD. The raw data are presented in Table S3 .

    Techniques Used: Imaging, Isolation, Expressing, Two Tailed Test

    Absence of RACK1 in astrocytes alters network population activity and neuronal responses to intense stimulation (A) Schematic of electrode positions used to record field excitatory postsynaptic potentials (fEPSPs) evoked by SC stimulation in the CA1 region of hippocampal slices. (B) Input-output curves for basal synaptic transmission. Left: representative recordings in RACK1 fl/fl mice (black) and RACK1 cKO mice before (pink) and after (blue) application of a Kir 4.1 antagonist (VU). Scale bars, 10 ms, 0.5 mV. Right: quantification of the fEPSP slope for different fiber volley amplitudes after SC stimulation. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 slices from 4 mice; p = 0.0087; RACK1 cKO: n = 5 slices from 5 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (C) Top: a representative recording of fEPSPs evoked by repetitive stimulation (10 Hz, 30 s) of CA1 SCs in RACK1 fl/fl mice under control conditions. Scale bars, 5 s, 0.2 mV. Bottom: enlarged view of fEPSPs evoked by the first 10 stimuli. Scale bars, 200 ms, 0.2 mV. (D) Changes in the fEPSP slope induced by 10-Hz stimulation relative to responses measured before the onset of stimulation (baseline responses) in RACK1 fl/fl mice (white filled dots) and in RACK1 cKO mice (pink-filled dots) before (black) and after (blue) application of VU. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 from 5 mice; RACK1 cKO: n = 6 slices from 4 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (E) Schematic (left) and picture (right) of a hippocampal slice placed on a multielectrode array (MEA). Scale bar, 200 μm. (F) Representative MEA recordings of burst activity induced in hippocampal slices of RACK1 fl/fl (black) and RACK1 cKO (pink) mice by incubation in Mg 2+ -free ACSF containing 6 mM KCl. The expanded recordings of the bursts (surrounded by gray rectangles) are shown on the right. Scale bars, 10 s (left)/200 ms (right), 50 μV. (G) Quantification of burst frequency (top) and burst duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices. The data are quoted as the mean ± SD. n = 15 slices from 5 mice for RACK1 fl/fl and n = 18 slices from 6 mice for RACK1 cKO; unpaired t test. (H) Representative MEA recordings of hippocampal bursting activity in RACK1 fl/fl (top) and RACK1 cKO (bottom) slices in control (Ct) and during 25-min treatment with the Kir4.1 blocker VU0134992 (VU). The corresponding time-frequency plots are shown under the traces. Scale bar, 20 s/2 min, 50 μV. (I) Quantification of VU’s effect on burst frequency (top) and duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices (RACK1 fl/fl: n = 15 slices from 5 mice for burst frequency and duration, respectively; RACK1 cKO: n = 18 slices from 6 mice for burst frequency and duration; paired t test. The data are quoted as the mean ± SD. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in <xref ref-type=Table S3 . " title="... control (Ct) and during 25-min treatment with the Kir4.1 blocker VU0134992 (VU). The corresponding time-frequency plots are ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Absence of RACK1 in astrocytes alters network population activity and neuronal responses to intense stimulation (A) Schematic of electrode positions used to record field excitatory postsynaptic potentials (fEPSPs) evoked by SC stimulation in the CA1 region of hippocampal slices. (B) Input-output curves for basal synaptic transmission. Left: representative recordings in RACK1 fl/fl mice (black) and RACK1 cKO mice before (pink) and after (blue) application of a Kir 4.1 antagonist (VU). Scale bars, 10 ms, 0.5 mV. Right: quantification of the fEPSP slope for different fiber volley amplitudes after SC stimulation. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 slices from 4 mice; p = 0.0087; RACK1 cKO: n = 5 slices from 5 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (C) Top: a representative recording of fEPSPs evoked by repetitive stimulation (10 Hz, 30 s) of CA1 SCs in RACK1 fl/fl mice under control conditions. Scale bars, 5 s, 0.2 mV. Bottom: enlarged view of fEPSPs evoked by the first 10 stimuli. Scale bars, 200 ms, 0.2 mV. (D) Changes in the fEPSP slope induced by 10-Hz stimulation relative to responses measured before the onset of stimulation (baseline responses) in RACK1 fl/fl mice (white filled dots) and in RACK1 cKO mice (pink-filled dots) before (black) and after (blue) application of VU. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 from 5 mice; RACK1 cKO: n = 6 slices from 4 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (E) Schematic (left) and picture (right) of a hippocampal slice placed on a multielectrode array (MEA). Scale bar, 200 μm. (F) Representative MEA recordings of burst activity induced in hippocampal slices of RACK1 fl/fl (black) and RACK1 cKO (pink) mice by incubation in Mg 2+ -free ACSF containing 6 mM KCl. The expanded recordings of the bursts (surrounded by gray rectangles) are shown on the right. Scale bars, 10 s (left)/200 ms (right), 50 μV. (G) Quantification of burst frequency (top) and burst duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices. The data are quoted as the mean ± SD. n = 15 slices from 5 mice for RACK1 fl/fl and n = 18 slices from 6 mice for RACK1 cKO; unpaired t test. (H) Representative MEA recordings of hippocampal bursting activity in RACK1 fl/fl (top) and RACK1 cKO (bottom) slices in control (Ct) and during 25-min treatment with the Kir4.1 blocker VU0134992 (VU). The corresponding time-frequency plots are shown under the traces. Scale bar, 20 s/2 min, 50 μV. (I) Quantification of VU’s effect on burst frequency (top) and duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices (RACK1 fl/fl: n = 15 slices from 5 mice for burst frequency and duration, respectively; RACK1 cKO: n = 18 slices from 6 mice for burst frequency and duration; paired t test. The data are quoted as the mean ± SD. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 .

    Techniques Used: Activity Assay, Transmission Assay, Incubation


    Figure Legend Snippet:

    Techniques Used: Transduction, Recombinant, Multiplex Assay, Mass Spectrometry, Software

    rabbit polyclonal kir4 1 antibody  (Alomone Labs)


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    Alomone Labs rabbit polyclonal kir4 1 antibody
    Rabbit Polyclonal Kir4 1 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit polyclonal anti kir4 1  (Alomone Labs)


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    Alomone Labs rabbit polyclonal anti kir4 1
    (A) Experimental design for (B)–(H). (B) Representative images of maximal projection and 3D reconstruction (IMARIS) of mG + astrocytes. (C and D) Surface area and enclosed volume of mG + astrocytes. Graphed as means of each astrocyte in Aldh1l1-CreER T2 :Sox2 fl/fl mice (left, n = 28 control, 43 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (E–H) Automatic tracing of mG + astrocyte processes by the filament tool of IMARIS and quantifications. Graphed as means of each astrocyte (left, n = 19 control, 17 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (I) Left, experimental design for (J)–(P); right, immunofluorescence of SOX2 and GFAP and quantification. n = 4 control, 4 Sox2 icKO. (J) Ctx astrocytes visualized by SR101 (arrows) for whole-cell recordings. (K) Voltage steps for astrocyte recording, from −180 to +20 mV with a step size of 10 mV. (L and M) Representative current tracing (L) and I-V curve (M) of one Sox2 -deficient and one Sox2 -intact astrocyte. (N1–N3) Quantification of cell capacitance (N1, n = 23 control, 18 Sox2 icKO), input resistance (N2, n = 20 control, 19 Sox2 icKO), and resting membrane potential (N3, n = 20 control, 17 Sox2 icKO) of Ctx astrocytes. (O) Western blot and quantification of <t>Kir4.1</t> in the brain. n = 4 control, 4 Sox2 icKO. (P) Representative images and quantification of Kir4.1 intensity in the cortex. n = 4 control, 4 Sox2 icKO. Error bars indicate means ± SEM. Unpaired two-tailed Student’s t test was used for statistically analyzing two groups of data. Please see for statistics. n, biological replicates. Scale bars, (B, E, I, J, and P) 20 μm.
    Rabbit Polyclonal Anti Kir4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "SOX2 is essential for astrocyte maturation and its deletion leads to hyperactive behavior in mice"

    Article Title: SOX2 is essential for astrocyte maturation and its deletion leads to hyperactive behavior in mice

    Journal: Cell reports

    doi: 10.1016/j.celrep.2022.111842

    (A) Experimental design for (B)–(H). (B) Representative images of maximal projection and 3D reconstruction (IMARIS) of mG + astrocytes. (C and D) Surface area and enclosed volume of mG + astrocytes. Graphed as means of each astrocyte in Aldh1l1-CreER T2 :Sox2 fl/fl mice (left, n = 28 control, 43 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (E–H) Automatic tracing of mG + astrocyte processes by the filament tool of IMARIS and quantifications. Graphed as means of each astrocyte (left, n = 19 control, 17 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (I) Left, experimental design for (J)–(P); right, immunofluorescence of SOX2 and GFAP and quantification. n = 4 control, 4 Sox2 icKO. (J) Ctx astrocytes visualized by SR101 (arrows) for whole-cell recordings. (K) Voltage steps for astrocyte recording, from −180 to +20 mV with a step size of 10 mV. (L and M) Representative current tracing (L) and I-V curve (M) of one Sox2 -deficient and one Sox2 -intact astrocyte. (N1–N3) Quantification of cell capacitance (N1, n = 23 control, 18 Sox2 icKO), input resistance (N2, n = 20 control, 19 Sox2 icKO), and resting membrane potential (N3, n = 20 control, 17 Sox2 icKO) of Ctx astrocytes. (O) Western blot and quantification of Kir4.1 in the brain. n = 4 control, 4 Sox2 icKO. (P) Representative images and quantification of Kir4.1 intensity in the cortex. n = 4 control, 4 Sox2 icKO. Error bars indicate means ± SEM. Unpaired two-tailed Student’s t test was used for statistically analyzing two groups of data. Please see for statistics. n, biological replicates. Scale bars, (B, E, I, J, and P) 20 μm.
    Figure Legend Snippet: (A) Experimental design for (B)–(H). (B) Representative images of maximal projection and 3D reconstruction (IMARIS) of mG + astrocytes. (C and D) Surface area and enclosed volume of mG + astrocytes. Graphed as means of each astrocyte in Aldh1l1-CreER T2 :Sox2 fl/fl mice (left, n = 28 control, 43 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (E–H) Automatic tracing of mG + astrocyte processes by the filament tool of IMARIS and quantifications. Graphed as means of each astrocyte (left, n = 19 control, 17 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (I) Left, experimental design for (J)–(P); right, immunofluorescence of SOX2 and GFAP and quantification. n = 4 control, 4 Sox2 icKO. (J) Ctx astrocytes visualized by SR101 (arrows) for whole-cell recordings. (K) Voltage steps for astrocyte recording, from −180 to +20 mV with a step size of 10 mV. (L and M) Representative current tracing (L) and I-V curve (M) of one Sox2 -deficient and one Sox2 -intact astrocyte. (N1–N3) Quantification of cell capacitance (N1, n = 23 control, 18 Sox2 icKO), input resistance (N2, n = 20 control, 19 Sox2 icKO), and resting membrane potential (N3, n = 20 control, 17 Sox2 icKO) of Ctx astrocytes. (O) Western blot and quantification of Kir4.1 in the brain. n = 4 control, 4 Sox2 icKO. (P) Representative images and quantification of Kir4.1 intensity in the cortex. n = 4 control, 4 Sox2 icKO. Error bars indicate means ± SEM. Unpaired two-tailed Student’s t test was used for statistically analyzing two groups of data. Please see for statistics. n, biological replicates. Scale bars, (B, E, I, J, and P) 20 μm.

    Techniques Used: Immunofluorescence, Western Blot, Two Tailed Test

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Protein Extraction, Recombinant, Electron Microscopy, Injection, Magnetic Cell Separation, Lysis, Bicinchoninic Acid Protein Assay, Western Blot, Glutamate Assay, Labeling, SYBR Green Assay, Chromatin Immunoprecipitation, Software, Activity Assay

    polyclonal rabbit antibody  (Alomone Labs)


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    Alomone Labs polyclonal rabbit antibody
    Polyclonal Rabbit Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit polyclonal  (Alomone Labs)


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    Alomone Labs rabbit polyclonal

    Rabbit Polyclonal, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Megalencephalic leukoencephalopathy with subcortical cysts is a developmental disorder of the gliovascular unit"

    Article Title: Megalencephalic leukoencephalopathy with subcortical cysts is a developmental disorder of the gliovascular unit

    Journal: eLife

    doi: 10.7554/eLife.71379


    Figure Legend Snippet:

    Techniques Used: Sequencing, Western Blot, Immunofluorescence, Transduction

    rabbit polyclonal  (Alomone Labs)


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    Alomone Labs rabbit polyclonal
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    rabbit polyclonal antibody against k ir 4 1  (Alomone Labs)


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    Alomone Labs rabbit polyclonal antibody against k ir 4 1
    List of primers used for the quantitative RT-PCR
    Rabbit Polyclonal Antibody Against K Ir 4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Targeted deletion of β1-syntrophin causes a loss of K ir 4.1 from Müller cell endfeet in mouse retina"

    Article Title: Targeted deletion of β1-syntrophin causes a loss of K ir 4.1 from Müller cell endfeet in mouse retina

    Journal: Glia

    doi: 10.1002/glia.23600


    Figure Legend Snippet: List of primers used for the quantitative RT-PCR

    Techniques Used:

    rabbit polyclonal anti body against kir4 1  (Alomone Labs)


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    Alomone Labs rabbit polyclonal anti body against kir4 1
    Rabbit Polyclonal Anti Body Against Kir4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit polyclonal  (Alomone Labs)


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    Alomone Labs rabbit polyclonal
    Rabbit Polyclonal, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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  • 96
    Alomone Labs rabbit polyclonal anti kir4 1
    (A) Experimental design for (B)–(H). (B) Representative images of maximal projection and 3D reconstruction (IMARIS) of mG + astrocytes. (C and D) Surface area and enclosed volume of mG + astrocytes. Graphed as means of each astrocyte in Aldh1l1-CreER T2 :Sox2 fl/fl mice (left, n = 28 control, 43 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (E–H) Automatic tracing of mG + astrocyte processes by the filament tool of IMARIS and quantifications. Graphed as means of each astrocyte (left, n = 19 control, 17 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (I) Left, experimental design for (J)–(P); right, immunofluorescence of SOX2 and GFAP and quantification. n = 4 control, 4 Sox2 icKO. (J) Ctx astrocytes visualized by SR101 (arrows) for whole-cell recordings. (K) Voltage steps for astrocyte recording, from −180 to +20 mV with a step size of 10 mV. (L and M) Representative current tracing (L) and I-V curve (M) of one Sox2 -deficient and one Sox2 -intact astrocyte. (N1–N3) Quantification of cell capacitance (N1, n = 23 control, 18 Sox2 icKO), input resistance (N2, n = 20 control, 19 Sox2 icKO), and resting membrane potential (N3, n = 20 control, 17 Sox2 icKO) of Ctx astrocytes. (O) Western blot and quantification of <t>Kir4.1</t> in the brain. n = 4 control, 4 Sox2 icKO. (P) Representative images and quantification of Kir4.1 intensity in the cortex. n = 4 control, 4 Sox2 icKO. Error bars indicate means ± SEM. Unpaired two-tailed Student’s t test was used for statistically analyzing two groups of data. Please see for statistics. n, biological replicates. Scale bars, (B, E, I, J, and P) 20 μm.
    Rabbit Polyclonal Anti Kir4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs rabbit polyclonal anti kir4 1 extracellular
    RACK1 KO in astrocytes leads to higher levels of <t>Kir4.1</t> in astrocyte somata and PAPs (A) Generation of a mouse line with RACK1 KO in astrocytes (RACK1 cKO). Shown is a schematic of the RACK1 fl/fl and Aldh1l1-Cre/ERT2 alleles. Deletion of exon 2 in Gnb2l1 (the gene coding for RACK1) is induced in astrocytes by tamoxifen injection; this results in a frameshift and premature termination of Gnb2l1 translation. Primers are indicated by red arrows. (B) PCR assays for Gnb2l1 KO in brain DNA from RACK1 fl/fl or Aldh1l1-CreERT2: RACK1 fl/fl tamoxifen-injected mice (RACK1 cKO). The 898-bp band corresponds to the floxed allele ( <xref ref-type=Table S4 ). The 672-bp band corresponds to the exon 2-deleted allele. (C) Confocal images of RACK1 immunofluorescence (red) in the hippocampus in RACK1 fl/fl and RACK1 cKO mice. Astrocytes are co-immunolabeled for GFAP (green). Some astrocytes are indicated by white arrowheads. Nuclei are stained with DAPI. The bottom panel gives a higher-magnification view of the boxed areas in the RACK1 fl/fl and RACK1 cKO images, which shows that RACK1 is specifically depleted in astrocytes ( ∗ ) and is still expressed by neurons (°). Scale bars, 20 μm. (D), Western blot detection and analysis of Kir4.1 and GLT-1 in protein extracts from whole brain, hippocampus, whole-brain synaptogliosomes, hippocampal synaptogliosomes, or whole-brain microvessels purified from RACK1 fl/fl or RACK1 cKO mice. The position of molecular weight markers is indicated on the right. Signals were normalized against that of stain-free membranes except for the experiment on purified microvessels, where histone 3 was used. The data are quoted as the mean ± SD (N = 5 samples per genotype, 1 mouse per sample); two-tailed unpaired t test or Mann-Whitney test. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 and Figure S2 . " width="250" height="auto" />
    Rabbit Polyclonal Anti Kir4 1 Extracellular, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Alomone Labs rabbit polyclonal kir4 1 antibody
    RACK1 KO in astrocytes leads to higher levels of <t>Kir4.1</t> in astrocyte somata and PAPs (A) Generation of a mouse line with RACK1 KO in astrocytes (RACK1 cKO). Shown is a schematic of the RACK1 fl/fl and Aldh1l1-Cre/ERT2 alleles. Deletion of exon 2 in Gnb2l1 (the gene coding for RACK1) is induced in astrocytes by tamoxifen injection; this results in a frameshift and premature termination of Gnb2l1 translation. Primers are indicated by red arrows. (B) PCR assays for Gnb2l1 KO in brain DNA from RACK1 fl/fl or Aldh1l1-CreERT2: RACK1 fl/fl tamoxifen-injected mice (RACK1 cKO). The 898-bp band corresponds to the floxed allele ( <xref ref-type=Table S4 ). The 672-bp band corresponds to the exon 2-deleted allele. (C) Confocal images of RACK1 immunofluorescence (red) in the hippocampus in RACK1 fl/fl and RACK1 cKO mice. Astrocytes are co-immunolabeled for GFAP (green). Some astrocytes are indicated by white arrowheads. Nuclei are stained with DAPI. The bottom panel gives a higher-magnification view of the boxed areas in the RACK1 fl/fl and RACK1 cKO images, which shows that RACK1 is specifically depleted in astrocytes ( ∗ ) and is still expressed by neurons (°). Scale bars, 20 μm. (D), Western blot detection and analysis of Kir4.1 and GLT-1 in protein extracts from whole brain, hippocampus, whole-brain synaptogliosomes, hippocampal synaptogliosomes, or whole-brain microvessels purified from RACK1 fl/fl or RACK1 cKO mice. The position of molecular weight markers is indicated on the right. Signals were normalized against that of stain-free membranes except for the experiment on purified microvessels, where histone 3 was used. The data are quoted as the mean ± SD (N = 5 samples per genotype, 1 mouse per sample); two-tailed unpaired t test or Mann-Whitney test. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 and Figure S2 . " width="250" height="auto" />
    Rabbit Polyclonal Kir4 1 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Alomone Labs polyclonal rabbit antibody
    RACK1 KO in astrocytes leads to higher levels of <t>Kir4.1</t> in astrocyte somata and PAPs (A) Generation of a mouse line with RACK1 KO in astrocytes (RACK1 cKO). Shown is a schematic of the RACK1 fl/fl and Aldh1l1-Cre/ERT2 alleles. Deletion of exon 2 in Gnb2l1 (the gene coding for RACK1) is induced in astrocytes by tamoxifen injection; this results in a frameshift and premature termination of Gnb2l1 translation. Primers are indicated by red arrows. (B) PCR assays for Gnb2l1 KO in brain DNA from RACK1 fl/fl or Aldh1l1-CreERT2: RACK1 fl/fl tamoxifen-injected mice (RACK1 cKO). The 898-bp band corresponds to the floxed allele ( <xref ref-type=Table S4 ). The 672-bp band corresponds to the exon 2-deleted allele. (C) Confocal images of RACK1 immunofluorescence (red) in the hippocampus in RACK1 fl/fl and RACK1 cKO mice. Astrocytes are co-immunolabeled for GFAP (green). Some astrocytes are indicated by white arrowheads. Nuclei are stained with DAPI. The bottom panel gives a higher-magnification view of the boxed areas in the RACK1 fl/fl and RACK1 cKO images, which shows that RACK1 is specifically depleted in astrocytes ( ∗ ) and is still expressed by neurons (°). Scale bars, 20 μm. (D), Western blot detection and analysis of Kir4.1 and GLT-1 in protein extracts from whole brain, hippocampus, whole-brain synaptogliosomes, hippocampal synaptogliosomes, or whole-brain microvessels purified from RACK1 fl/fl or RACK1 cKO mice. The position of molecular weight markers is indicated on the right. Signals were normalized against that of stain-free membranes except for the experiment on purified microvessels, where histone 3 was used. The data are quoted as the mean ± SD (N = 5 samples per genotype, 1 mouse per sample); two-tailed unpaired t test or Mann-Whitney test. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 and Figure S2 . " width="250" height="auto" />
    Polyclonal Rabbit Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs rabbit polyclonal

    Rabbit Polyclonal, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs rabbit polyclonal antibody against k ir 4 1
    List of primers used for the quantitative RT-PCR
    Rabbit Polyclonal Antibody Against K Ir 4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs rabbit polyclonal anti body against kir4 1
    List of primers used for the quantitative RT-PCR
    Rabbit Polyclonal Anti Body Against Kir4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    (A) Experimental design for (B)–(H). (B) Representative images of maximal projection and 3D reconstruction (IMARIS) of mG + astrocytes. (C and D) Surface area and enclosed volume of mG + astrocytes. Graphed as means of each astrocyte in Aldh1l1-CreER T2 :Sox2 fl/fl mice (left, n = 28 control, 43 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (E–H) Automatic tracing of mG + astrocyte processes by the filament tool of IMARIS and quantifications. Graphed as means of each astrocyte (left, n = 19 control, 17 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (I) Left, experimental design for (J)–(P); right, immunofluorescence of SOX2 and GFAP and quantification. n = 4 control, 4 Sox2 icKO. (J) Ctx astrocytes visualized by SR101 (arrows) for whole-cell recordings. (K) Voltage steps for astrocyte recording, from −180 to +20 mV with a step size of 10 mV. (L and M) Representative current tracing (L) and I-V curve (M) of one Sox2 -deficient and one Sox2 -intact astrocyte. (N1–N3) Quantification of cell capacitance (N1, n = 23 control, 18 Sox2 icKO), input resistance (N2, n = 20 control, 19 Sox2 icKO), and resting membrane potential (N3, n = 20 control, 17 Sox2 icKO) of Ctx astrocytes. (O) Western blot and quantification of Kir4.1 in the brain. n = 4 control, 4 Sox2 icKO. (P) Representative images and quantification of Kir4.1 intensity in the cortex. n = 4 control, 4 Sox2 icKO. Error bars indicate means ± SEM. Unpaired two-tailed Student’s t test was used for statistically analyzing two groups of data. Please see for statistics. n, biological replicates. Scale bars, (B, E, I, J, and P) 20 μm.

    Journal: Cell reports

    Article Title: SOX2 is essential for astrocyte maturation and its deletion leads to hyperactive behavior in mice

    doi: 10.1016/j.celrep.2022.111842

    Figure Lengend Snippet: (A) Experimental design for (B)–(H). (B) Representative images of maximal projection and 3D reconstruction (IMARIS) of mG + astrocytes. (C and D) Surface area and enclosed volume of mG + astrocytes. Graphed as means of each astrocyte in Aldh1l1-CreER T2 :Sox2 fl/fl mice (left, n = 28 control, 43 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (E–H) Automatic tracing of mG + astrocyte processes by the filament tool of IMARIS and quantifications. Graphed as means of each astrocyte (left, n = 19 control, 17 Sox2 icKO) and mouse (right, n = 3 control, 4 Sox2 icKO). (I) Left, experimental design for (J)–(P); right, immunofluorescence of SOX2 and GFAP and quantification. n = 4 control, 4 Sox2 icKO. (J) Ctx astrocytes visualized by SR101 (arrows) for whole-cell recordings. (K) Voltage steps for astrocyte recording, from −180 to +20 mV with a step size of 10 mV. (L and M) Representative current tracing (L) and I-V curve (M) of one Sox2 -deficient and one Sox2 -intact astrocyte. (N1–N3) Quantification of cell capacitance (N1, n = 23 control, 18 Sox2 icKO), input resistance (N2, n = 20 control, 19 Sox2 icKO), and resting membrane potential (N3, n = 20 control, 17 Sox2 icKO) of Ctx astrocytes. (O) Western blot and quantification of Kir4.1 in the brain. n = 4 control, 4 Sox2 icKO. (P) Representative images and quantification of Kir4.1 intensity in the cortex. n = 4 control, 4 Sox2 icKO. Error bars indicate means ± SEM. Unpaired two-tailed Student’s t test was used for statistically analyzing two groups of data. Please see for statistics. n, biological replicates. Scale bars, (B, E, I, J, and P) 20 μm.

    Article Snippet: Rabbit polyclonal anti-Kir4.1 , Alomone labs , Cat# APC-035; RRID: AB_2040120.

    Techniques: Immunofluorescence, Western Blot, Two Tailed Test

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: SOX2 is essential for astrocyte maturation and its deletion leads to hyperactive behavior in mice

    doi: 10.1016/j.celrep.2022.111842

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Rabbit polyclonal anti-Kir4.1 , Alomone labs , Cat# APC-035; RRID: AB_2040120.

    Techniques: Protein Extraction, Recombinant, Electron Microscopy, Injection, Magnetic Cell Separation, Lysis, Bicinchoninic Acid Protein Assay, Western Blot, Glutamate Assay, Labeling, SYBR Green Assay, Chromatin Immunoprecipitation, Software, Activity Assay

    RACK1 KO in astrocytes leads to higher levels of Kir4.1 in astrocyte somata and PAPs (A) Generation of a mouse line with RACK1 KO in astrocytes (RACK1 cKO). Shown is a schematic of the RACK1 fl/fl and Aldh1l1-Cre/ERT2 alleles. Deletion of exon 2 in Gnb2l1 (the gene coding for RACK1) is induced in astrocytes by tamoxifen injection; this results in a frameshift and premature termination of Gnb2l1 translation. Primers are indicated by red arrows. (B) PCR assays for Gnb2l1 KO in brain DNA from RACK1 fl/fl or Aldh1l1-CreERT2: RACK1 fl/fl tamoxifen-injected mice (RACK1 cKO). The 898-bp band corresponds to the floxed allele ( <xref ref-type=Table S4 ). The 672-bp band corresponds to the exon 2-deleted allele. (C) Confocal images of RACK1 immunofluorescence (red) in the hippocampus in RACK1 fl/fl and RACK1 cKO mice. Astrocytes are co-immunolabeled for GFAP (green). Some astrocytes are indicated by white arrowheads. Nuclei are stained with DAPI. The bottom panel gives a higher-magnification view of the boxed areas in the RACK1 fl/fl and RACK1 cKO images, which shows that RACK1 is specifically depleted in astrocytes ( ∗ ) and is still expressed by neurons (°). Scale bars, 20 μm. (D), Western blot detection and analysis of Kir4.1 and GLT-1 in protein extracts from whole brain, hippocampus, whole-brain synaptogliosomes, hippocampal synaptogliosomes, or whole-brain microvessels purified from RACK1 fl/fl or RACK1 cKO mice. The position of molecular weight markers is indicated on the right. Signals were normalized against that of stain-free membranes except for the experiment on purified microvessels, where histone 3 was used. The data are quoted as the mean ± SD (N = 5 samples per genotype, 1 mouse per sample); two-tailed unpaired t test or Mann-Whitney test. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 and Figure S2 . " width="100%" height="100%">

    Journal: Cell Reports

    Article Title: The ribosome-associated protein RACK1 represses Kir4.1 translation in astrocytes and influences neuronal activity

    doi: 10.1016/j.celrep.2023.112456

    Figure Lengend Snippet: RACK1 KO in astrocytes leads to higher levels of Kir4.1 in astrocyte somata and PAPs (A) Generation of a mouse line with RACK1 KO in astrocytes (RACK1 cKO). Shown is a schematic of the RACK1 fl/fl and Aldh1l1-Cre/ERT2 alleles. Deletion of exon 2 in Gnb2l1 (the gene coding for RACK1) is induced in astrocytes by tamoxifen injection; this results in a frameshift and premature termination of Gnb2l1 translation. Primers are indicated by red arrows. (B) PCR assays for Gnb2l1 KO in brain DNA from RACK1 fl/fl or Aldh1l1-CreERT2: RACK1 fl/fl tamoxifen-injected mice (RACK1 cKO). The 898-bp band corresponds to the floxed allele ( Table S4 ). The 672-bp band corresponds to the exon 2-deleted allele. (C) Confocal images of RACK1 immunofluorescence (red) in the hippocampus in RACK1 fl/fl and RACK1 cKO mice. Astrocytes are co-immunolabeled for GFAP (green). Some astrocytes are indicated by white arrowheads. Nuclei are stained with DAPI. The bottom panel gives a higher-magnification view of the boxed areas in the RACK1 fl/fl and RACK1 cKO images, which shows that RACK1 is specifically depleted in astrocytes ( ∗ ) and is still expressed by neurons (°). Scale bars, 20 μm. (D), Western blot detection and analysis of Kir4.1 and GLT-1 in protein extracts from whole brain, hippocampus, whole-brain synaptogliosomes, hippocampal synaptogliosomes, or whole-brain microvessels purified from RACK1 fl/fl or RACK1 cKO mice. The position of molecular weight markers is indicated on the right. Signals were normalized against that of stain-free membranes except for the experiment on purified microvessels, where histone 3 was used. The data are quoted as the mean ± SD (N = 5 samples per genotype, 1 mouse per sample); two-tailed unpaired t test or Mann-Whitney test. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 and Figure S2 .

    Article Snippet: Rabbit polyclonal anti-KIR4.1 extracellular , Alomone labs , Cat#APC-165; RRID: AB_2341043.

    Techniques: Injection, Immunofluorescence, Immunolabeling, Staining, Western Blot, Purification, Molecular Weight, Two Tailed Test, MANN-WHITNEY

    Absence of RACK1 in astrocytes leads to higher Kir4.1-mediated astrocytic inward K + currents and cell volumes (A) Schematic of electrode positions used to record astrocyte whole-cell currents evoked by Schaffer collateral (SC) stimulation in the CA1 region of hippocampal slices. (B) Left: representative traces of astrocytic whole-cell currents induced by 150-ms voltage steps (from −200 mV to +100 mV, 10-mV steps; black traces at the bottom) in RACK1 fl/fl and RACK1 cKO mice before (black and pink) and after (blue) application of a KIR 4.1 antagonist (VU). Scale bars, 50 ms, 5 nA. Right: current-voltage (I-V) plot in RACK1 fl/fl (white filled dots) and in RACK1 cKO (pink-filled dots) mice before (black) and after (blue) application of VU (N = 7 and 10 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons) The data are quoted as the mean ± SD. (C) Left: representative astrocytic Kir4.1 (VU-sensitive) currents induced by SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (black) and RACK1 cKO (pink) mice. Scale bars, 200 ms, 50 pA. Right: quantification of astrocytic Kir4.1 current peak amplitude after each stimulus during SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (white filled dots) and RACK1 cKO (pink filled dots) mice (N = 6 and 5 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons). The data are quoted as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (D) Illustration of the imaging method with a representative raw confocal image of an isolated RACK1 fl/fl CA1 astrocyte expressing tdTomato. (E–G) Imaris analysis: filament tracing (E), convex hull volume (F), and a 3D Sholl analysis (G). (H) Mean territory volume and filament length of RACK1 fl/fl and RACK1 cKO astrocytes. Shown is a histogram of the data, presented as the mean ± SD (N = 4 mice per genotype, 45 astrocytes); two-tailed t test. The data are quoted as the mean ± SD. (I) A Sholl analysis of the ramification complexity of RACK1 fl/fl and RACK1 cKO astrocytes. Two-way analysis of variance. ∗ p < 0.05, ∗∗ p < 0.01. The data are quoted as the mean ± SD. The raw data are presented in <xref ref-type=Table S3 . " width="100%" height="100%">

    Journal: Cell Reports

    Article Title: The ribosome-associated protein RACK1 represses Kir4.1 translation in astrocytes and influences neuronal activity

    doi: 10.1016/j.celrep.2023.112456

    Figure Lengend Snippet: Absence of RACK1 in astrocytes leads to higher Kir4.1-mediated astrocytic inward K + currents and cell volumes (A) Schematic of electrode positions used to record astrocyte whole-cell currents evoked by Schaffer collateral (SC) stimulation in the CA1 region of hippocampal slices. (B) Left: representative traces of astrocytic whole-cell currents induced by 150-ms voltage steps (from −200 mV to +100 mV, 10-mV steps; black traces at the bottom) in RACK1 fl/fl and RACK1 cKO mice before (black and pink) and after (blue) application of a KIR 4.1 antagonist (VU). Scale bars, 50 ms, 5 nA. Right: current-voltage (I-V) plot in RACK1 fl/fl (white filled dots) and in RACK1 cKO (pink-filled dots) mice before (black) and after (blue) application of VU (N = 7 and 10 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons) The data are quoted as the mean ± SD. (C) Left: representative astrocytic Kir4.1 (VU-sensitive) currents induced by SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (black) and RACK1 cKO (pink) mice. Scale bars, 200 ms, 50 pA. Right: quantification of astrocytic Kir4.1 current peak amplitude after each stimulus during SC stimulation (10 Hz, 1 s) in RACK1 fl/fl (white filled dots) and RACK1 cKO (pink filled dots) mice (N = 6 and 5 astrocytes for RACK1 fl/fl and RACK1 cKO mice, respectively; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons). The data are quoted as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (D) Illustration of the imaging method with a representative raw confocal image of an isolated RACK1 fl/fl CA1 astrocyte expressing tdTomato. (E–G) Imaris analysis: filament tracing (E), convex hull volume (F), and a 3D Sholl analysis (G). (H) Mean territory volume and filament length of RACK1 fl/fl and RACK1 cKO astrocytes. Shown is a histogram of the data, presented as the mean ± SD (N = 4 mice per genotype, 45 astrocytes); two-tailed t test. The data are quoted as the mean ± SD. (I) A Sholl analysis of the ramification complexity of RACK1 fl/fl and RACK1 cKO astrocytes. Two-way analysis of variance. ∗ p < 0.05, ∗∗ p < 0.01. The data are quoted as the mean ± SD. The raw data are presented in Table S3 .

    Article Snippet: Rabbit polyclonal anti-KIR4.1 extracellular , Alomone labs , Cat#APC-165; RRID: AB_2341043.

    Techniques: Imaging, Isolation, Expressing, Two Tailed Test

    Absence of RACK1 in astrocytes alters network population activity and neuronal responses to intense stimulation (A) Schematic of electrode positions used to record field excitatory postsynaptic potentials (fEPSPs) evoked by SC stimulation in the CA1 region of hippocampal slices. (B) Input-output curves for basal synaptic transmission. Left: representative recordings in RACK1 fl/fl mice (black) and RACK1 cKO mice before (pink) and after (blue) application of a Kir 4.1 antagonist (VU). Scale bars, 10 ms, 0.5 mV. Right: quantification of the fEPSP slope for different fiber volley amplitudes after SC stimulation. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 slices from 4 mice; p = 0.0087; RACK1 cKO: n = 5 slices from 5 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (C) Top: a representative recording of fEPSPs evoked by repetitive stimulation (10 Hz, 30 s) of CA1 SCs in RACK1 fl/fl mice under control conditions. Scale bars, 5 s, 0.2 mV. Bottom: enlarged view of fEPSPs evoked by the first 10 stimuli. Scale bars, 200 ms, 0.2 mV. (D) Changes in the fEPSP slope induced by 10-Hz stimulation relative to responses measured before the onset of stimulation (baseline responses) in RACK1 fl/fl mice (white filled dots) and in RACK1 cKO mice (pink-filled dots) before (black) and after (blue) application of VU. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 from 5 mice; RACK1 cKO: n = 6 slices from 4 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (E) Schematic (left) and picture (right) of a hippocampal slice placed on a multielectrode array (MEA). Scale bar, 200 μm. (F) Representative MEA recordings of burst activity induced in hippocampal slices of RACK1 fl/fl (black) and RACK1 cKO (pink) mice by incubation in Mg 2+ -free ACSF containing 6 mM KCl. The expanded recordings of the bursts (surrounded by gray rectangles) are shown on the right. Scale bars, 10 s (left)/200 ms (right), 50 μV. (G) Quantification of burst frequency (top) and burst duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices. The data are quoted as the mean ± SD. n = 15 slices from 5 mice for RACK1 fl/fl and n = 18 slices from 6 mice for RACK1 cKO; unpaired t test. (H) Representative MEA recordings of hippocampal bursting activity in RACK1 fl/fl (top) and RACK1 cKO (bottom) slices in control (Ct) and during 25-min treatment with the Kir4.1 blocker VU0134992 (VU). The corresponding time-frequency plots are shown under the traces. Scale bar, 20 s/2 min, 50 μV. (I) Quantification of VU’s effect on burst frequency (top) and duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices (RACK1 fl/fl: n = 15 slices from 5 mice for burst frequency and duration, respectively; RACK1 cKO: n = 18 slices from 6 mice for burst frequency and duration; paired t test. The data are quoted as the mean ± SD. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in <xref ref-type=Table S3 . " width="100%" height="100%">

    Journal: Cell Reports

    Article Title: The ribosome-associated protein RACK1 represses Kir4.1 translation in astrocytes and influences neuronal activity

    doi: 10.1016/j.celrep.2023.112456

    Figure Lengend Snippet: Absence of RACK1 in astrocytes alters network population activity and neuronal responses to intense stimulation (A) Schematic of electrode positions used to record field excitatory postsynaptic potentials (fEPSPs) evoked by SC stimulation in the CA1 region of hippocampal slices. (B) Input-output curves for basal synaptic transmission. Left: representative recordings in RACK1 fl/fl mice (black) and RACK1 cKO mice before (pink) and after (blue) application of a Kir 4.1 antagonist (VU). Scale bars, 10 ms, 0.5 mV. Right: quantification of the fEPSP slope for different fiber volley amplitudes after SC stimulation. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 slices from 4 mice; p = 0.0087; RACK1 cKO: n = 5 slices from 5 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (C) Top: a representative recording of fEPSPs evoked by repetitive stimulation (10 Hz, 30 s) of CA1 SCs in RACK1 fl/fl mice under control conditions. Scale bars, 5 s, 0.2 mV. Bottom: enlarged view of fEPSPs evoked by the first 10 stimuli. Scale bars, 200 ms, 0.2 mV. (D) Changes in the fEPSP slope induced by 10-Hz stimulation relative to responses measured before the onset of stimulation (baseline responses) in RACK1 fl/fl mice (white filled dots) and in RACK1 cKO mice (pink-filled dots) before (black) and after (blue) application of VU. The data are quoted as the mean ± SD. RACK1 fl/fl: n = 5 from 5 mice; RACK1 cKO: n = 6 slices from 4 mice; repeated-measures two-way ANOVA with Sidak’s correction for multiple comparisons. (E) Schematic (left) and picture (right) of a hippocampal slice placed on a multielectrode array (MEA). Scale bar, 200 μm. (F) Representative MEA recordings of burst activity induced in hippocampal slices of RACK1 fl/fl (black) and RACK1 cKO (pink) mice by incubation in Mg 2+ -free ACSF containing 6 mM KCl. The expanded recordings of the bursts (surrounded by gray rectangles) are shown on the right. Scale bars, 10 s (left)/200 ms (right), 50 μV. (G) Quantification of burst frequency (top) and burst duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices. The data are quoted as the mean ± SD. n = 15 slices from 5 mice for RACK1 fl/fl and n = 18 slices from 6 mice for RACK1 cKO; unpaired t test. (H) Representative MEA recordings of hippocampal bursting activity in RACK1 fl/fl (top) and RACK1 cKO (bottom) slices in control (Ct) and during 25-min treatment with the Kir4.1 blocker VU0134992 (VU). The corresponding time-frequency plots are shown under the traces. Scale bar, 20 s/2 min, 50 μV. (I) Quantification of VU’s effect on burst frequency (top) and duration (bottom) in RACK1 fl/fl (white) and RACK1 cKO (pink) hippocampal slices (RACK1 fl/fl: n = 15 slices from 5 mice for burst frequency and duration, respectively; RACK1 cKO: n = 18 slices from 6 mice for burst frequency and duration; paired t test. The data are quoted as the mean ± SD. ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. The raw data are presented in Table S3 .

    Article Snippet: Rabbit polyclonal anti-KIR4.1 extracellular , Alomone labs , Cat#APC-165; RRID: AB_2341043.

    Techniques: Activity Assay, Transmission Assay, Incubation

    Journal: Cell Reports

    Article Title: The ribosome-associated protein RACK1 represses Kir4.1 translation in astrocytes and influences neuronal activity

    doi: 10.1016/j.celrep.2023.112456

    Figure Lengend Snippet:

    Article Snippet: Rabbit polyclonal anti-KIR4.1 extracellular , Alomone labs , Cat#APC-165; RRID: AB_2341043.

    Techniques: Transduction, Recombinant, Multiplex Assay, Mass Spectrometry, Software

    Journal: eLife

    Article Title: Megalencephalic leukoencephalopathy with subcortical cysts is a developmental disorder of the gliovascular unit

    doi: 10.7554/eLife.71379

    Figure Lengend Snippet:

    Article Snippet: Antibody , Kir 4.1 extracellular (rabbit polyclonal) , Alomone Labs , APC-165 , Immunofluorescence (1:200).

    Techniques: Sequencing, Western Blot, Immunofluorescence, Transduction

    Journal: Glia

    Article Title: Targeted deletion of β1-syntrophin causes a loss of K ir 4.1 from Müller cell endfeet in mouse retina

    doi: 10.1002/glia.23600

    Figure Lengend Snippet: List of primers used for the quantitative RT-PCR

    Article Snippet: Primary antibodies were: i) affinity-purified rabbit polyclonal antibody against K ir 4.1 (1:200 dilution; Alomone Labs; Cat# APC-035, RRID:AB_2040120); ii) affinity-purified rabbit polyclonal antibody against β1-syn (Syn248; 1:100 dilution; ( Peters et al., 1997 )).

    Techniques: