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Covalab Inc p eb2 s222
Induced Neurons Model Molecular Consequences of CDKL5 Variants but Display NoTranscriptional, Morphological or Functional Changes. A) Normalized log fold gene expression for iN samples. B) Representative western blots for <t>EB2</t> and phosphorylation of EB2 S221 with TUJ1/Beta-3-tubulin as a loading control. C) Paired t-test of phosphorylation of EB2 fold change relative to EB2 between isogenic control and CDD patient-derived neurons (N=3 for 2 donors). D) Pearson correlation coefficients for gene expression for the 5000 most variable genes between all samples from iNs. E) Volcano plot of differentially expressed genes between pooled isogenic control and CDD patient-derived samples (N=6 per genotype). F) Representative images of iN neurites immunolabeled with TUJ1 and MAP2 in both patient and isogenic control lines. A paired t-test was performed for changes in neurite length normalized to the cell body area (μm/μm 2 ) for G) MAP2 immunofluorescence (N=3 batches). H) Synchrony index and I) weighted mean firing rate for INs during the first 30 days of maturation post-plating (N=4 differentiation batches, 2 donors). Error bars represent the standard error of the mean.
P Eb2 S222, supplied by Covalab Inc, 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/p eb2 s222/product/Covalab Inc
Average 86 stars, based on 1 article reviews
p eb2 s222 - by Bioz Stars, 2025-02
86/100 stars

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1) Product Images from "Excitatory Cortical Neurons from CDKL5 Deficiency Disorder Patient-Derived Organoids Show Early Hyperexcitability Not Identified in Neurogenin2 Induced Neurons"

Article Title: Excitatory Cortical Neurons from CDKL5 Deficiency Disorder Patient-Derived Organoids Show Early Hyperexcitability Not Identified in Neurogenin2 Induced Neurons

Journal: bioRxiv

doi: 10.1101/2024.11.11.622878

Induced Neurons Model Molecular Consequences of CDKL5 Variants but Display NoTranscriptional, Morphological or Functional Changes. A) Normalized log fold gene expression for iN samples. B) Representative western blots for EB2 and phosphorylation of EB2 S221 with TUJ1/Beta-3-tubulin as a loading control. C) Paired t-test of phosphorylation of EB2 fold change relative to EB2 between isogenic control and CDD patient-derived neurons (N=3 for 2 donors). D) Pearson correlation coefficients for gene expression for the 5000 most variable genes between all samples from iNs. E) Volcano plot of differentially expressed genes between pooled isogenic control and CDD patient-derived samples (N=6 per genotype). F) Representative images of iN neurites immunolabeled with TUJ1 and MAP2 in both patient and isogenic control lines. A paired t-test was performed for changes in neurite length normalized to the cell body area (μm/μm 2 ) for G) MAP2 immunofluorescence (N=3 batches). H) Synchrony index and I) weighted mean firing rate for INs during the first 30 days of maturation post-plating (N=4 differentiation batches, 2 donors). Error bars represent the standard error of the mean.
Figure Legend Snippet: Induced Neurons Model Molecular Consequences of CDKL5 Variants but Display NoTranscriptional, Morphological or Functional Changes. A) Normalized log fold gene expression for iN samples. B) Representative western blots for EB2 and phosphorylation of EB2 S221 with TUJ1/Beta-3-tubulin as a loading control. C) Paired t-test of phosphorylation of EB2 fold change relative to EB2 between isogenic control and CDD patient-derived neurons (N=3 for 2 donors). D) Pearson correlation coefficients for gene expression for the 5000 most variable genes between all samples from iNs. E) Volcano plot of differentially expressed genes between pooled isogenic control and CDD patient-derived samples (N=6 per genotype). F) Representative images of iN neurites immunolabeled with TUJ1 and MAP2 in both patient and isogenic control lines. A paired t-test was performed for changes in neurite length normalized to the cell body area (μm/μm 2 ) for G) MAP2 immunofluorescence (N=3 batches). H) Synchrony index and I) weighted mean firing rate for INs during the first 30 days of maturation post-plating (N=4 differentiation batches, 2 donors). Error bars represent the standard error of the mean.

Techniques Used: Functional Assay, Expressing, Western Blot, Control, Derivative Assay, Immunolabeling, Immunofluorescence

No Altered mTOR Signaling in iNs. A) Representative Western blots for phosphorylation of different mTOR-signaling associated proteins. pEB2 and EB2 as shown in for reference. B) Quantification of normalized protein abundance for each tested protein (N=3). Each shape represents a differentiation batch.
Figure Legend Snippet: No Altered mTOR Signaling in iNs. A) Representative Western blots for phosphorylation of different mTOR-signaling associated proteins. pEB2 and EB2 as shown in for reference. B) Quantification of normalized protein abundance for each tested protein (N=3). Each shape represents a differentiation batch.

Techniques Used: Western Blot

Cortical ACTX Neurons with CDKL5 Variants Have Reduced pEB2 and Decreased Expression of CDKL5 . A) Representative western blot of EB2 and pEB2 from ACTX neurons. B) Fold change quantification of western blots for phosphorylation of EB2 relative to EB2 across 3 donors (N = 4 donors). C) Pearson correlations across ACTX neuronal differentiations. D) Differentially expressed genes between pooled isogenic controls and CDD proband-derived neurons (N=7 sample per genotype across 3 donors). E) log2 fold change of CDKL5 and HS3ST1 in ACTX neurons of patient cell lines relative to isogenic controls (N=3, paired t-test). EIF2A is the house-keeping gene.
Figure Legend Snippet: Cortical ACTX Neurons with CDKL5 Variants Have Reduced pEB2 and Decreased Expression of CDKL5 . A) Representative western blot of EB2 and pEB2 from ACTX neurons. B) Fold change quantification of western blots for phosphorylation of EB2 relative to EB2 across 3 donors (N = 4 donors). C) Pearson correlations across ACTX neuronal differentiations. D) Differentially expressed genes between pooled isogenic controls and CDD proband-derived neurons (N=7 sample per genotype across 3 donors). E) log2 fold change of CDKL5 and HS3ST1 in ACTX neurons of patient cell lines relative to isogenic controls (N=3, paired t-test). EIF2A is the house-keeping gene.

Techniques Used: Expressing, Western Blot, Derivative Assay



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Covalab Inc p eb2 s222
Induced Neurons Model Molecular Consequences of CDKL5 Variants but Display NoTranscriptional, Morphological or Functional Changes. A) Normalized log fold gene expression for iN samples. B) Representative western blots for <t>EB2</t> and phosphorylation of EB2 S221 with TUJ1/Beta-3-tubulin as a loading control. C) Paired t-test of phosphorylation of EB2 fold change relative to EB2 between isogenic control and CDD patient-derived neurons (N=3 for 2 donors). D) Pearson correlation coefficients for gene expression for the 5000 most variable genes between all samples from iNs. E) Volcano plot of differentially expressed genes between pooled isogenic control and CDD patient-derived samples (N=6 per genotype). F) Representative images of iN neurites immunolabeled with TUJ1 and MAP2 in both patient and isogenic control lines. A paired t-test was performed for changes in neurite length normalized to the cell body area (μm/μm 2 ) for G) MAP2 immunofluorescence (N=3 batches). H) Synchrony index and I) weighted mean firing rate for INs during the first 30 days of maturation post-plating (N=4 differentiation batches, 2 donors). Error bars represent the standard error of the mean.
P Eb2 S222, supplied by Covalab Inc, 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/p eb2 s222/product/Covalab Inc
Average 86 stars, based on 1 article reviews
p eb2 s222 - by Bioz Stars, 2025-02
86/100 stars
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Induced Neurons Model Molecular Consequences of CDKL5 Variants but Display NoTranscriptional, Morphological or Functional Changes. A) Normalized log fold gene expression for iN samples. B) Representative western blots for EB2 and phosphorylation of EB2 S221 with TUJ1/Beta-3-tubulin as a loading control. C) Paired t-test of phosphorylation of EB2 fold change relative to EB2 between isogenic control and CDD patient-derived neurons (N=3 for 2 donors). D) Pearson correlation coefficients for gene expression for the 5000 most variable genes between all samples from iNs. E) Volcano plot of differentially expressed genes between pooled isogenic control and CDD patient-derived samples (N=6 per genotype). F) Representative images of iN neurites immunolabeled with TUJ1 and MAP2 in both patient and isogenic control lines. A paired t-test was performed for changes in neurite length normalized to the cell body area (μm/μm 2 ) for G) MAP2 immunofluorescence (N=3 batches). H) Synchrony index and I) weighted mean firing rate for INs during the first 30 days of maturation post-plating (N=4 differentiation batches, 2 donors). Error bars represent the standard error of the mean.

Journal: bioRxiv

Article Title: Excitatory Cortical Neurons from CDKL5 Deficiency Disorder Patient-Derived Organoids Show Early Hyperexcitability Not Identified in Neurogenin2 Induced Neurons

doi: 10.1101/2024.11.11.622878

Figure Lengend Snippet: Induced Neurons Model Molecular Consequences of CDKL5 Variants but Display NoTranscriptional, Morphological or Functional Changes. A) Normalized log fold gene expression for iN samples. B) Representative western blots for EB2 and phosphorylation of EB2 S221 with TUJ1/Beta-3-tubulin as a loading control. C) Paired t-test of phosphorylation of EB2 fold change relative to EB2 between isogenic control and CDD patient-derived neurons (N=3 for 2 donors). D) Pearson correlation coefficients for gene expression for the 5000 most variable genes between all samples from iNs. E) Volcano plot of differentially expressed genes between pooled isogenic control and CDD patient-derived samples (N=6 per genotype). F) Representative images of iN neurites immunolabeled with TUJ1 and MAP2 in both patient and isogenic control lines. A paired t-test was performed for changes in neurite length normalized to the cell body area (μm/μm 2 ) for G) MAP2 immunofluorescence (N=3 batches). H) Synchrony index and I) weighted mean firing rate for INs during the first 30 days of maturation post-plating (N=4 differentiation batches, 2 donors). Error bars represent the standard error of the mean.

Article Snippet: Membranes were then incubated for one hour at room temperature with EB2 1:500 (Abcam ab234843), 1:500 p-EB2 S222 (CovalAb pab01032-P) Signaling Technologies #5364), 1:500 S6 (Santa Cruz #sc-74459), 1:500 p-AKT (Sigma #05-1003), 1:500 AKT (Cell Signaling Technologies #4691S), 1:500 p-GSK (Cell Signaling Technologies #5558), 1:500 GSK (Cell Signaling Technologies #5676S) 1:4000 Acetylated tubulin (Sigma #T7451), 1:4000 TUJ1 (Sigma #T8660), 1:4000 Tyrosinated tubulin (Millipore #MAB 1864), 1:2000 GAPDH (Invitrogen #AM4300), washed by TBS-T, incubated for one hour at room temperature with secondary antibodies at 1:10000 (Alexa Fluor488, ThermoFisher #A28175; Alexa Fluor568, ThermoFisher #A-11011) and imaged using the Odyssey imaging system (LI-COR).

Techniques: Functional Assay, Expressing, Western Blot, Control, Derivative Assay, Immunolabeling, Immunofluorescence

No Altered mTOR Signaling in iNs. A) Representative Western blots for phosphorylation of different mTOR-signaling associated proteins. pEB2 and EB2 as shown in for reference. B) Quantification of normalized protein abundance for each tested protein (N=3). Each shape represents a differentiation batch.

Journal: bioRxiv

Article Title: Excitatory Cortical Neurons from CDKL5 Deficiency Disorder Patient-Derived Organoids Show Early Hyperexcitability Not Identified in Neurogenin2 Induced Neurons

doi: 10.1101/2024.11.11.622878

Figure Lengend Snippet: No Altered mTOR Signaling in iNs. A) Representative Western blots for phosphorylation of different mTOR-signaling associated proteins. pEB2 and EB2 as shown in for reference. B) Quantification of normalized protein abundance for each tested protein (N=3). Each shape represents a differentiation batch.

Article Snippet: Membranes were then incubated for one hour at room temperature with EB2 1:500 (Abcam ab234843), 1:500 p-EB2 S222 (CovalAb pab01032-P) Signaling Technologies #5364), 1:500 S6 (Santa Cruz #sc-74459), 1:500 p-AKT (Sigma #05-1003), 1:500 AKT (Cell Signaling Technologies #4691S), 1:500 p-GSK (Cell Signaling Technologies #5558), 1:500 GSK (Cell Signaling Technologies #5676S) 1:4000 Acetylated tubulin (Sigma #T7451), 1:4000 TUJ1 (Sigma #T8660), 1:4000 Tyrosinated tubulin (Millipore #MAB 1864), 1:2000 GAPDH (Invitrogen #AM4300), washed by TBS-T, incubated for one hour at room temperature with secondary antibodies at 1:10000 (Alexa Fluor488, ThermoFisher #A28175; Alexa Fluor568, ThermoFisher #A-11011) and imaged using the Odyssey imaging system (LI-COR).

Techniques: Western Blot

Cortical ACTX Neurons with CDKL5 Variants Have Reduced pEB2 and Decreased Expression of CDKL5 . A) Representative western blot of EB2 and pEB2 from ACTX neurons. B) Fold change quantification of western blots for phosphorylation of EB2 relative to EB2 across 3 donors (N = 4 donors). C) Pearson correlations across ACTX neuronal differentiations. D) Differentially expressed genes between pooled isogenic controls and CDD proband-derived neurons (N=7 sample per genotype across 3 donors). E) log2 fold change of CDKL5 and HS3ST1 in ACTX neurons of patient cell lines relative to isogenic controls (N=3, paired t-test). EIF2A is the house-keeping gene.

Journal: bioRxiv

Article Title: Excitatory Cortical Neurons from CDKL5 Deficiency Disorder Patient-Derived Organoids Show Early Hyperexcitability Not Identified in Neurogenin2 Induced Neurons

doi: 10.1101/2024.11.11.622878

Figure Lengend Snippet: Cortical ACTX Neurons with CDKL5 Variants Have Reduced pEB2 and Decreased Expression of CDKL5 . A) Representative western blot of EB2 and pEB2 from ACTX neurons. B) Fold change quantification of western blots for phosphorylation of EB2 relative to EB2 across 3 donors (N = 4 donors). C) Pearson correlations across ACTX neuronal differentiations. D) Differentially expressed genes between pooled isogenic controls and CDD proband-derived neurons (N=7 sample per genotype across 3 donors). E) log2 fold change of CDKL5 and HS3ST1 in ACTX neurons of patient cell lines relative to isogenic controls (N=3, paired t-test). EIF2A is the house-keeping gene.

Article Snippet: Membranes were then incubated for one hour at room temperature with EB2 1:500 (Abcam ab234843), 1:500 p-EB2 S222 (CovalAb pab01032-P) Signaling Technologies #5364), 1:500 S6 (Santa Cruz #sc-74459), 1:500 p-AKT (Sigma #05-1003), 1:500 AKT (Cell Signaling Technologies #4691S), 1:500 p-GSK (Cell Signaling Technologies #5558), 1:500 GSK (Cell Signaling Technologies #5676S) 1:4000 Acetylated tubulin (Sigma #T7451), 1:4000 TUJ1 (Sigma #T8660), 1:4000 Tyrosinated tubulin (Millipore #MAB 1864), 1:2000 GAPDH (Invitrogen #AM4300), washed by TBS-T, incubated for one hour at room temperature with secondary antibodies at 1:10000 (Alexa Fluor488, ThermoFisher #A28175; Alexa Fluor568, ThermoFisher #A-11011) and imaged using the Odyssey imaging system (LI-COR).

Techniques: Expressing, Western Blot, Derivative Assay