Journal:

Article Title: Caveolin 1 Is Required for the Activation of Endothelial Nitric Oxide Synthase in Response to 17?-Estradiol

doi: 10.1210/me.2010-0043

Figure Lengend Snippet: Decreasing caveolin 1 expression attenuates 17β-estradiol-mediated activation of eNOS. PAEC were transfected with either a caveolin 1 siRNA or a scrambled siRNA (as a control). Immunoblot analysis demonstrates a significant decrease in caveolin 1 protein levels in caveolin 1 siRNA-transfected cells (A and B; *, P < 0.05 vs. scrambled siRNA). Plasma membrane fractions were isolated and subjected to immunoblotting with either an antibody against ERα or eNOS. Decreasing caveolin 1 expression results in a significant reduction in ERα localization to the plasma membrane (C and D; *, P < 0.05 vs. scrambled siRNA) although total ERα levels are unchanged (C). The siRNA-mediated knockdown of caveolin 1 does not alter eNOS localized to the plasma membrane (E and F), Immunoblotting with the plasma membrane marker 5′-nucleotidase was also carried out to normalize for protein loading of the plasma membrane fractions (C and E). The siRNA-mediated knockdown of caveolin 1 significantly inhibits 17β-estradiol-stimulated (100 nm, 30 min) eNOS phosphorylation at serine 1177 (G and H) and NO generation (I). The siRNA-mediated knockdown of caveolin 1 does not inhibit the increase in NO generation in response to stimulation by shear stress (20 dyn/cm2, 15 min; panel J) or VEGF (100 ng/ml, 15min; panel K). Data are mean ± sem, n = 3–6. *, P < 0.05 vs. no 17β-estradiol; †, P < 0.05 vs. 17β-estradiol-treated scrambled siRNA.

Article Snippet: Generation and analysis of a caveolin 1 peptide array A library of overlapping peptides (12 mers), each shifted by five amino acids across the entire sequence of caveolin 1, was immobilized on cellulose membranes (Intavis Bioanalytical Instrruments AG, Cologne, Germany) and probed with 100 ng/ml recombinant ERα.

Techniques: Expressing, Activation Assay, Transfection, Western Blot, Isolation, Marker

Journal:

Article Title: Caveolin 1 Is Required for the Activation of Endothelial Nitric Oxide Synthase in Response to 17?-Estradiol

doi: 10.1210/me.2010-0043

Figure Lengend Snippet: 17β-Estradiol stimulates caveolin 1 phosphorylation through the activation of pp60Src. PAEC were exposed to pp60Src inhibitor, PP2 (10 μm, 30 min), before 17β-estradiol (100 nm, 30 min). 17β-Estradiol significantly increases phospho-Tyr14 caveolin 1, and PP2 inhibits this phosphorylation although total caveolin 1 levels are unchanged (A and B). 17β-Estradiol also produces significantly higher eNOS phosphorylation at serine 1177 compared with untreated cells, and pretreatment with the ER antagonist, ICI 182 780 (10 μm, 30 min) or PP2 significantly attenuates eNOS phosphorylation at serine 1177 (C and D). 17β-Estradiol treatment also induces a significant increase in NO generation, and pretreatment with ICI 182,780 or PP2 results in significant reduction in NO production (E). Data are mean ± sem n = 3–4. *, P < 0.05 vs. untreated; †, P < 0.05 vs. 17β-estradiol treated.

Article Snippet: Generation and analysis of a caveolin 1 peptide array A library of overlapping peptides (12 mers), each shifted by five amino acids across the entire sequence of caveolin 1, was immobilized on cellulose membranes (Intavis Bioanalytical Instrruments AG, Cologne, Germany) and probed with 100 ng/ml recombinant ERα.

Techniques: Activation Assay

Journal:

Article Title: Caveolin 1 Is Required for the Activation of Endothelial Nitric Oxide Synthase in Response to 17?-Estradiol

doi: 10.1210/me.2010-0043

Figure Lengend Snippet: Caveolin 1 phosphorylation is not required for the localization of ERα to the plasma membrane. Immunoblot analysis of PAEC transduced with adenoviral constructs for GFP (AdGFP), wild-type caveolin 1 (AdWTCav) or a tyrosine 14-deficient mutant (AdY14FCav) indicates significant overexpression of AdWTCav and AdY14Cav (A and B). There is significantly higher ERα in the plasma membrane in both AdWTCav- and AdY14Cav-transduced cells (C and D). AdY14Cav overexpression did not alter eNOS localization to the plasma membrane (E and F). Immunoblotting with the plasma membrane marker 5′-nucleotidase was also carried out to normalize for protein loading of the plasma membrane fractions (C and E). Immunofluorescence analysis of cells transduced with AdGFP, AdWTCav-1, or AdY14FCav-1 were then transfected with an ERα-GFP construct. The plasma membrane of these cells was labeled with Alexa Fluor 594 WGA (red). The extent of membrane localization of ERα was then determined by measuring the intensity of yellow fluorescence (overlap of red fluorescence of Alexa Fluor 594 WGA and green fluorescence of ERα-GFP (G); the white bar in each individual panel represents 30 μm). The intensity of yellow fluorescence is significantly higher in cells transduced with either AdWTCav or AdY14Cav (H). Data are mean ± sem n = 3. *, P < 0.05 vs. AdGFP-transduced cells.

Article Snippet: Generation and analysis of a caveolin 1 peptide array A library of overlapping peptides (12 mers), each shifted by five amino acids across the entire sequence of caveolin 1, was immobilized on cellulose membranes (Intavis Bioanalytical Instrruments AG, Cologne, Germany) and probed with 100 ng/ml recombinant ERα.

Techniques: Western Blot, Transduction, Construct, Mutagenesis, Over Expression, Marker, Immunofluorescence, Transfection, Labeling, Fluorescence

Journal:

Article Title: Caveolin 1 Is Required for the Activation of Endothelial Nitric Oxide Synthase in Response to 17?-Estradiol

doi: 10.1210/me.2010-0043

Figure Lengend Snippet: Caveolin 1 phosphorylation enhances the interaction of ERα with PI3 kinase. PAEC were transduced with AdGFP, AdWTCav-1, or AdY14FCav-1, exposed or not to 17β-estradiol, and the interaction of ERα with caveolin 1 (A and C) or PI3 kinase (A and B) was determined. 17β-Estradiol exposure causes a dissociation of the ERα-caveolin 1 complex (A and C). This dissociation was enhanced in cells overexpressing wild-type caveolin 1 and attenuated in cells overexpressing Y14F caveolin 1 (A and C). 17β-Estradiol also significantly increases the interaction of ERα with PI3 kinase (A and B). Transduction with AdWTCav potentiates the 17β-estradiol-mediated interaction (A and B) whereas AdY14FCav attenuates this effect (A and B). Data are mean ± sem; n = 3. *, P < 0.05 vs. untreated cells; †, P < 0.05 vs. AdGFP with 17β-estradiol. IB, Immunoblotting; IP, immunoprecipitation.

Article Snippet: Generation and analysis of a caveolin 1 peptide array A library of overlapping peptides (12 mers), each shifted by five amino acids across the entire sequence of caveolin 1, was immobilized on cellulose membranes (Intavis Bioanalytical Instrruments AG, Cologne, Germany) and probed with 100 ng/ml recombinant ERα.

Techniques: Transduction, Western Blot, Immunoprecipitation

Journal:

Article Title: Caveolin 1 Is Required for the Activation of Endothelial Nitric Oxide Synthase in Response to 17?-Estradiol

doi: 10.1210/me.2010-0043

Figure Lengend Snippet: Caveolin 1 phosphorylation enhances Akt activation and NO signaling in response to 17β-estradiol. PAEC were transduced with AdGFP, AdWTCav-1, or AdY14FCav-1, exposed or not to 17β-estradiol, and the effect on Akt activity (A and B), eNOS phosphorylation at serine1177 (C and D), and NO generation (E) was determined. The 17β-estradiol-mediated increase in Akt activity (A and B), eNOS phosphorylation at serine 1177 (C and D), and NO generation (E) is enhanced in cells overexpressing wild-type caveolin 1 and attenuated in cells overexpressing Y14F caveolin 1. The overexpression of wild-type caveolin 1 (F and G) or AdY14F (H and I) did not significantly attenuate the increase in NO generation in response to stimulation by shear stress (20 dyn/cm2, 15 min) or VEGF (100 ng/ml, 15min). Data are mean ± sem; n =3–6. *, P < 0.05 vs. untreated; †, P < 0.05 vs. AdGFP with 17β-estradiol.

Article Snippet: Generation and analysis of a caveolin 1 peptide array A library of overlapping peptides (12 mers), each shifted by five amino acids across the entire sequence of caveolin 1, was immobilized on cellulose membranes (Intavis Bioanalytical Instrruments AG, Cologne, Germany) and probed with 100 ng/ml recombinant ERα.

Techniques: Activation Assay, Transduction, Activity Assay, Over Expression

Journal:

Article Title: Caveolin 1 Is Required for the Activation of Endothelial Nitric Oxide Synthase in Response to 17?-Estradiol

doi: 10.1210/me.2010-0043

Figure Lengend Snippet: The scaffold domain of caveolin 1 is required for ERα trafficking to the plasma membrane but not for protein-protein interaction. Immunoblot analysis of PAEC transduced with adenoviral constructs for GFP (AdGFP), wild-type caveolin 1 (AdWTCav), or a caveolin 1 mutant lacking the scaffold domain (AdCavΔ) indicates significant overexpression of AdWTCav and AdCavΔ (A and B). There is significantly higher ERα (C and D) and caveolin 1 (C and E) in the plasma membrane in the AdWTCav-transduced cells. However, both ERα (C and D) and caveolin 1 (C and E) are significantly decreased in the AdCavΔ-transduced cells (C and E). Immunoblotting (IB) with the plasma membrane marker 5′-nucleotidase was also carried out to normalize for protein loading of the plasma membrane fractions (C). Immunoprecipitation (IP) of caveolin 1 from whole-cell extracts followed by IB with ERα identified a significant increase in ERα bound to caveolin 1 in both AdWTCav- and AdCavΔ-transduced cells (F and G). Data are mean ± sem; n = 3. *, P < 0.05 vs. AdGFP-transduced cells.

Article Snippet: Generation and analysis of a caveolin 1 peptide array A library of overlapping peptides (12 mers), each shifted by five amino acids across the entire sequence of caveolin 1, was immobilized on cellulose membranes (Intavis Bioanalytical Instrruments AG, Cologne, Germany) and probed with 100 ng/ml recombinant ERα.

Techniques: Western Blot, Transduction, Construct, Mutagenesis, Over Expression, Marker, Immunoprecipitation

Journal:

Article Title: Caveolin 1 Is Required for the Activation of Endothelial Nitric Oxide Synthase in Response to 17?-Estradiol

doi: 10.1210/me.2010-0043

Figure Lengend Snippet: Amino acids 62–73 of caveolin 1 span the ERα interaction region. Probing a peptide array representing the entire sequence of human caveolin 1 with recombinant human ERα (100 ng/ml) identified a single highly reactive peptide spanning amino acids 62–73 (shown by arrow in panel A). TAT-Cav62–73, along with a scrambled version containing a similar amino acid composition (TAT-Cav62–73scr), was synthesized as a fusion with the N-terminal TAT protein transduction domain from the HIV (B). PAEC were exposed to each peptide (100 ng/ml). Immunopreciptation (IP) analyses demonstrate that TAT-Cav62–73 significantly attenuates the association of ERα with caveolin 1 (C and D). Western blot analysis also demonstrates that TAT-Cav62–73 significantly attenuates ERα levels in the plasma membrane without altering total ERα levels (E and F; *, P < 0.05 vs. TAT-Cav62–73scr). Immunoblotting (IB) with the plasma membrane marker 5′-nucleotidase was also carried out to normalize for protein loading of the plasma membrane fractions (E). TAT-Cav62–73 and TAT-Cav62–73scr peptide-treated cells were transfected with an ERα-GFP construct and the plasma membrane labeled with Alexa Fluor 594 WGA (red). The extent of plasma membrane localization of ERα was determined by measuring the intensity of yellow fluorescence (overlap of red fluorescence of Alexa Fluor 594 WGA, and green fluorescence of ERα-GFP). TAT-Cav62–73 significantly attenuates the yellow fluorescence (G and H, the white bar in each panel represents 30 μm). TAT-Cav62–73 did not alter eNOS localization to the plasma membrane (I and J). n = 3; *, P < 0.05 vs. TAT-Cav62–73scr.

Article Snippet: Generation and analysis of a caveolin 1 peptide array A library of overlapping peptides (12 mers), each shifted by five amino acids across the entire sequence of caveolin 1, was immobilized on cellulose membranes (Intavis Bioanalytical Instrruments AG, Cologne, Germany) and probed with 100 ng/ml recombinant ERα.

Techniques: Peptide Microarray, Sequencing, Recombinant, Synthesized, Transduction, Western Blot, Marker, Transfection, Construct, Labeling, Fluorescence

Journal:

Article Title: Caveolin 1 Is Required for the Activation of Endothelial Nitric Oxide Synthase in Response to 17?-Estradiol

doi: 10.1210/me.2010-0043

Figure Lengend Snippet: Disrupting ERα-caveolin 1 interactions attenuates the interaction of ERα with PI3 kinase and inhibits downstream NO signaling. TAT-Cav62–73 significantly attenuates the 17β-estradiol-mediated association of ERα with PI3 kinase (A and B), the increase in eNOS phosphorylation at serine 1177 (C and D), and the increase in NO generation (E). TAT-Cav62–73 did inhibit the increase in NO generation in response to stimulation by shear stress (20 dyn/cm2, 15 min; panel F) or VEGF (100 ng/ml, 15min; panel G). Data are mean ± sem; n = 3–6. *, P < 0.05 vs. untreated; †, P < 0.05 vs. TAT-Cav62–73scr with 17β-estradiol.

Article Snippet: Generation and analysis of a caveolin 1 peptide array A library of overlapping peptides (12 mers), each shifted by five amino acids across the entire sequence of caveolin 1, was immobilized on cellulose membranes (Intavis Bioanalytical Instrruments AG, Cologne, Germany) and probed with 100 ng/ml recombinant ERα.

Techniques:

Journal: The Journal of Physiological Sciences : JPS

Article Title: Transient receptor potential cation 3 channel regulates melanoma proliferation and migration

doi: 10.1007/s12576-016-0480-1

Figure Lengend Snippet: TRPC3 expression and SOCE in melanoma. a Representative images of immunohistochemical staining of HE, MART1, and TRPC3 in a melanoma primary tissue microarray (stage II) (original magnification, b ×200). The calibration bars represent 200 µm. b mRNA expression in various melanoma cell lines. SK-Mel-2 is a skin metastasis melanoma cell line with NRAS mutation, SK-Mel-24 is a lymph node metastasis cell line with BRAFV600E mutation, SK-Mel-187 is also a lymph node metastasis cell line, C8161 is a metastasis cell line with wild-type BRAF, and HEMA-LP is a skin melanocyte cell line. TRPC3 mRNA is widely expressed in human melanoma cell lines, independently of BRAF mutation. c Immunoprecipitation for TRPC3 and STIM1 was performed. d Cytosolic Ca2+ level in C8161 cells is shown as mean ± SD (n = 8–9). SOCE was examined in the presence or absence of DMSO (vehicle control 1 µM) or Pyr3 (1 µM) in C8161. Pyr3 inhibited SOCE in C8161 cells. Pyr3 was added after the addition of Ca2+. The Ca2+ signal was immediately decreased, indicating that Pyr3 inhibits SOCE in melanoma cells. Data in each panel are averages of eight or nine cells

Article Snippet: C8161 cells treated with DMSO or Pyr3 (10 μM) for 15 min was subjected to protein phosphorylation microarray assay using a commercial kit (Cancer Signaling Phospho-Antibody Array; Full Moon BioSystems, Inc.).

Techniques: Expressing, Immunohistochemical staining, Staining, Microarray, Mutagenesis, Immunoprecipitation, Control

Journal: The Journal of Physiological Sciences : JPS

Article Title: Transient receptor potential cation 3 channel regulates melanoma proliferation and migration

doi: 10.1007/s12576-016-0480-1

Figure Lengend Snippet: Pyr3 inhibits phosphorylation of STAT5 and Akt. a Representative images of Akt phosphorylation are shown. Densitometric analyses (bar graph) of Western blots showed that phosphorylation of Akt was inhibited by TRPC3 inhibitor, Pyr3 (n = 4, **p < 0.01, ns not significant). b Protein phosphorylation microarray analysis in the presence of Pyr3. C8161 cells were incubated with DMSO (vehicle control) or Pyr3 (10 μM) for 15 min. The Y-axis shows the signal ratio of phosphorylated to non-phosphorylated protein in the presence of Pyr3 as a percentage of that of the DMSO control. c Representative images of STAT5 phosphorylation. Densitometric analyses (bar graph) of Western blots showed that phosphorylation of STAT5 was inhibited by Pyr3 (n = 4, *p < 0.05, ns not significant)

Article Snippet: C8161 cells treated with DMSO or Pyr3 (10 μM) for 15 min was subjected to protein phosphorylation microarray assay using a commercial kit (Cancer Signaling Phospho-Antibody Array; Full Moon BioSystems, Inc.).

Techniques: Phospho-proteomics, Western Blot, Microarray, Incubation, Control

Journal: Clinical and Translational Science

Article Title: Immunostimulatory Effects of Mesenchymal Stem Cell‐Derived Neurons: Implications for Stem Cell Therapy in Allogeneic Transplantations

doi: 10.1111/j.1752-8062.2008.00018.x

Figure Lengend Snippet: Baseline cytokine production in mscs and PBmcs . Cell‐free media from uninduced MSCs and unstimulated PBMCs were analyzed for cytokine production using protein arrays. Each experiment was performed four times, each with cells from a different donor. The densitometric scans were normalized to 10, and the normalized alues are presented as the mean ± SD.

Article Snippet: Microarray for cytokines Cell‐free media from MLR reactions with uninduced and induced (8‐ and 12‐day) MSCs were studied for cytokine expressions with human cytokine protein array III (Ray Biotech, Norcross, GA, USA), as described.

Techniques:

Journal: Clinical and Translational Science

Article Title: Immunostimulatory Effects of Mesenchymal Stem Cell‐Derived Neurons: Implications for Stem Cell Therapy in Allogeneic Transplantations

doi: 10.1111/j.1752-8062.2008.00018.x

Figure Lengend Snippet: Cytokine produce in MLRs. Cell‐free media from MLR with uninduced (day 0) and induced MSCs (day 8 and day 12) were analyzed on cytokine arrays. Parallel studies with media from PBMC or MSCs cultured alone are shown in Table 1. Each experiment was performed four times, each with cells from a different donor. The densitometric scans were normalized to 10, and the normalized values are presented as the mean ± SD.

Article Snippet: Microarray for cytokines Cell‐free media from MLR reactions with uninduced and induced (8‐ and 12‐day) MSCs were studied for cytokine expressions with human cytokine protein array III (Ray Biotech, Norcross, GA, USA), as described.

Techniques: Cell Culture

Journal: Biomedicines

Article Title: The Enhanced Expression of ZWILCH Predicts Poor Survival of Adrenocortical Carcinoma Patients.

doi: 10.3390/biomedicines11041233

Figure Lengend Snippet: Figure 1. An overall survival area plot showing different estimates of the causal effect of the ZWILCH expression on survival using different Cox proportional hazards regression models (A). The contour plot illustrates the probability of patient survival over time according to ZWILCH gene expression level in ACC patients from the TCGA database (B).

Article Snippet: Slides were immersed in Target Retrieval Solution, Citrate pH 6.1 (#H-3300-250, Vector, Stuttgart, Germany), and heated in a microwave for 5 min. After that, they were cooled down to room temperature (RT) for 20 min. After endogenous peroxidase activity blocking and incubation with 2.5% normal horse serum, the sections were incubated with anti-ZWILCH polyclonal rabbit antibody (#14281-1-AP, Proteintech, Rosemont, IL, USA) with 1:1000 concentration at 4 ◦C overnight.

Techniques: Expressing, Gene Expression

Journal: Biomedicines

Article Title: The Enhanced Expression of ZWILCH Predicts Poor Survival of Adrenocortical Carcinoma Patients.

doi: 10.3390/biomedicines11041233

Figure Lengend Snippet: Figure 5. General transcriptome profile in groups of 10 patients with the highest expression of the ZWILCH gene (up ZWILCH) compared to 10 patients with the lowest expression of the ZWILCH gene (down ZWILCH) (data from GSE10927). Each dot on the graph represents the mean value of the gene expression level of ten patients belonging to a particular group. The cut-off criteria for identifying differentially expressed genes were established as |fold change| = 1.5 and p-value = 0.05, and are represented by orange dotted lines. Genes above the cut-off lines were categorized as up−regulated (green dots) or down−regulated (red dots). The total number of up−and down−regulated genes can be found in the top right and top left corners, respectively. The plots also indicate the symbols for the seven most differentially expressed genes.

Article Snippet: Slides were immersed in Target Retrieval Solution, Citrate pH 6.1 (#H-3300-250, Vector, Stuttgart, Germany), and heated in a microwave for 5 min. After that, they were cooled down to room temperature (RT) for 20 min. After endogenous peroxidase activity blocking and incubation with 2.5% normal horse serum, the sections were incubated with anti-ZWILCH polyclonal rabbit antibody (#14281-1-AP, Proteintech, Rosemont, IL, USA) with 1:1000 concentration at 4 ◦C overnight.

Techniques: Expressing, Gene Expression

Journal: Biomedicines

Article Title: The Enhanced Expression of ZWILCH Predicts Poor Survival of Adrenocortical Carcinoma Patients.

doi: 10.3390/biomedicines11041233

Figure Lengend Snippet: Figure 8. Gene set enrichment analysis (GSEA) from the comparison of patients with low and high ZWILCH expression, based on GSE10927. Clustering of enriched gene sets into common functional groups, each cluster is marked with a different color (A). The barplot with ten of the most activated and inhibited gene terms according to the normalized enrichment score values (B). Detailed enrichment plots for the five most inhibited and the five most activated gene sets, showing the profile of the running NES score and the positions of the genes on the rank−ordered list (C,D).

Article Snippet: Slides were immersed in Target Retrieval Solution, Citrate pH 6.1 (#H-3300-250, Vector, Stuttgart, Germany), and heated in a microwave for 5 min. After that, they were cooled down to room temperature (RT) for 20 min. After endogenous peroxidase activity blocking and incubation with 2.5% normal horse serum, the sections were incubated with anti-ZWILCH polyclonal rabbit antibody (#14281-1-AP, Proteintech, Rosemont, IL, USA) with 1:1000 concentration at 4 ◦C overnight.

Techniques: Comparison, Expressing, Functional Assay

Journal: Biomedicines

Article Title: The Enhanced Expression of ZWILCH Predicts Poor Survival of Adrenocortical Carcinoma Patients.

doi: 10.3390/biomedicines11041233

Figure Lengend Snippet: Figure 10. The expression of ZWILCH protein in human adrenal gland disease spectrum (adrenal cancer progression) tissue microarray (TMA) slide. The general profile of immunohistochemical staining presents localization of ZWILCH protein (A). TMA map shows the relevant types of adrenal cancer progression marked in an appropriate color (B). Densitometric analysis of ZWILCH gene expression in the tissue array side of the adrenal gland disease spectrum. The boxplot displays each group with its median and interquartile range (IQR) (C). Individual patient densitometric data were added to the corresponding boxplots and represented as dots. The Kruskal–Wallis (KW) test was used to compare groups, followed by the Dunn post hoc test. Differences between groups were marked with asterisks: ** for p < 0.01. Letter annotation was used to indicate statistically significant differences (p < 0.05) between compared groups. Paired comparisons with post hoc Dunn p-values are also shown in the table.

Article Snippet: Slides were immersed in Target Retrieval Solution, Citrate pH 6.1 (#H-3300-250, Vector, Stuttgart, Germany), and heated in a microwave for 5 min. After that, they were cooled down to room temperature (RT) for 20 min. After endogenous peroxidase activity blocking and incubation with 2.5% normal horse serum, the sections were incubated with anti-ZWILCH polyclonal rabbit antibody (#14281-1-AP, Proteintech, Rosemont, IL, USA) with 1:1000 concentration at 4 ◦C overnight.

Techniques: Expressing, Microarray, Immunohistochemical staining, Staining, Gene Expression

Journal: Biomedicines

Article Title: The Enhanced Expression of ZWILCH Predicts Poor Survival of Adrenocortical Carcinoma Patients.

doi: 10.3390/biomedicines11041233

Figure Lengend Snippet: Figure 11. Representative immunostaining of adrenal gland disease spectrum (adrenal cancer pro- gression) TMA slide. Brown staining (B,C,E,F,H,I,K,L,N,O) indicates ZWILCH protein (red arrows), located typically cellular with hematoxylin counterstain (nucleus). The negative control of adrenal gland tissue (A,D,G,J,L,M). Original magnification was 100× (A,B,D,E,G,H,M,N), 200× (J,L,K), and 400× (C,F,I,L,O).

Article Snippet: Slides were immersed in Target Retrieval Solution, Citrate pH 6.1 (#H-3300-250, Vector, Stuttgart, Germany), and heated in a microwave for 5 min. After that, they were cooled down to room temperature (RT) for 20 min. After endogenous peroxidase activity blocking and incubation with 2.5% normal horse serum, the sections were incubated with anti-ZWILCH polyclonal rabbit antibody (#14281-1-AP, Proteintech, Rosemont, IL, USA) with 1:1000 concentration at 4 ◦C overnight.

Techniques: Immunostaining, Staining, Negative Control

Journal: Stem cell research

Article Title: Generation and characterization of human induced pluripotent stem cells (iPSCs) from three male and three female patients with CDKL5 Deficiency Disorder (CDD)

doi: 10.1016/j.scr.2021.102276

Figure Lengend Snippet:

Article Snippet: Pluripotency Marker , Rabbit anti-OCT4 , 1:100 , Abcam Cat# ab19857, RRID: AB_445175.

Techniques: Derivative Assay, Sequencing, Modification

Journal: Stem cell research

Article Title: Generation and characterization of human induced pluripotent stem cells (iPSCs) from three male and three female patients with CDKL5 Deficiency Disorder (CDD)

doi: 10.1016/j.scr.2021.102276

Figure Lengend Snippet: Characterization and validation

Article Snippet: Pluripotency Marker , Rabbit anti-OCT4 , 1:100 , Abcam Cat# ab19857, RRID: AB_445175.

Techniques: Immunocytochemistry, Staining, Expressing, Microarray, Marker, Mutagenesis, Sequencing, Southern Blot

Journal: Stem cell research

Article Title: Generation and characterization of human induced pluripotent stem cells (iPSCs) from three male and three female patients with CDKL5 Deficiency Disorder (CDD)

doi: 10.1016/j.scr.2021.102276

Figure Lengend Snippet: Reagents details

Article Snippet: Pluripotency Marker , Rabbit anti-OCT4 , 1:100 , Abcam Cat# ab19857, RRID: AB_445175.

Techniques: Immunocytochemistry, Marker, Expressing, Mutagenesis, Sequencing