nicd  (Cell Signaling Technology Inc)

Journal: Pancreas

Article Title: Knockdown of Lymphoid Enhancer-binding Factor 1 Inhibits Pancreatic Adenocarcinoma Growth and Neoangiogenesis by Curbing Notch1 and Nuclear Factor Kappa B Signaling Pathways

doi: 10.1097/MPA.0000000000002562

Figure Lengend Snippet: Knockdown of LEF1 interferes with PAAD cell malignant progression and neovascularization by inhibiting Notch1 and NF-κB signaling pathways. A–F, Immunofluorescence experiments and fluorescence intensity analysis: the relative fluorescence level of Notch1 and P65 decreased in AsPC-1/LEF1 and BxPC-3/LEF1. G–M, WB assay results and quantitative analysis: the total Notch1, NICD, Hes1, Hey1, and Jagged1 protein levels and the p-P65/P65 ratio were decreased in AsPC-1/LEF1 and BxPC-3/LEF1 (n = 3, * P <0.05).

Article Snippet: The first antibodies were as follows: LEF1 (ab137872, 1:1000, Abcam), proliferating cell nuclear antigen (PCNA, ab29, 1:1000, Abcam), MMP-2 (4022S, 1:1000, CST), MMP-9 (3852S, 1:1000, CST), BAX (2772S, 1:1000, CST), Bcl-2 (15071S, 1:1000, CST), Cleaved caspase-3 (9661S, 1:1000, CST), Cleaved caspase-9 (9509S, 1:1000, CST), vascular endothelial growth factor-A (VEGFA, ab214424, 1:1000, Abcam), total Notch1 (ab52627, 1:1000, Abcam), NICD (4147, 1:1000, CST), Hes1 (ab71559, 1:1000, Abcam), Hey1 (ab235173, 1:1000, Abcam), Jagged1 (ab300561, 1:1000, Abcam), P65 (ab32536, 1:1000, Abcam), p-P65 (3033S, 1:1000, CST), GAPDH (5174S, 1:5000, CST), HRP-anti-Rabbit (A0362, 1:1000, Beyotime), HRP-anti-Mouse (A0350, 1:1000, Beyotime).

Techniques: Knockdown, Protein-Protein interactions, Immunofluorescence, Fluorescence

Journal: Pancreas

Article Title: Knockdown of Lymphoid Enhancer-binding Factor 1 Inhibits Pancreatic Adenocarcinoma Growth and Neoangiogenesis by Curbing Notch1 and Nuclear Factor Kappa B Signaling Pathways

doi: 10.1097/MPA.0000000000002562

Figure Lengend Snippet: Knockdown of LEF1 enhances immune response in mice. A and B, Immunofluorescence results and percentage of positive cells: the fluorescence intensity of PD-L1 in tumor tissues of sh-LEF1 group decreased. C and D, HE staining results and necrotic cell count showed that the proportion of cell necrosis in sh-LEF1 tumor tissue was decreased. E–H, ELISA results and content statistics showed that knockdown of LEF1 downregulated the contents of TGF-β1, IL-10, PD-L1, and CCL2 in sh-LEF1 tumor tissues. I–O, WB assay results and quantitative analysis showed decreased levels of total Notch1, NICD, Hes1, Hey1, and Jagged1 proteins in tumor tissues, and a decreased p-P65/P65 ratio (n = 6, * P <0.05).

Article Snippet: The first antibodies were as follows: LEF1 (ab137872, 1:1000, Abcam), proliferating cell nuclear antigen (PCNA, ab29, 1:1000, Abcam), MMP-2 (4022S, 1:1000, CST), MMP-9 (3852S, 1:1000, CST), BAX (2772S, 1:1000, CST), Bcl-2 (15071S, 1:1000, CST), Cleaved caspase-3 (9661S, 1:1000, CST), Cleaved caspase-9 (9509S, 1:1000, CST), vascular endothelial growth factor-A (VEGFA, ab214424, 1:1000, Abcam), total Notch1 (ab52627, 1:1000, Abcam), NICD (4147, 1:1000, CST), Hes1 (ab71559, 1:1000, Abcam), Hey1 (ab235173, 1:1000, Abcam), Jagged1 (ab300561, 1:1000, Abcam), P65 (ab32536, 1:1000, Abcam), p-P65 (3033S, 1:1000, CST), GAPDH (5174S, 1:5000, CST), HRP-anti-Rabbit (A0362, 1:1000, Beyotime), HRP-anti-Mouse (A0350, 1:1000, Beyotime).

Techniques: Knockdown, Immunofluorescence, Fluorescence, Staining, Cell Characterization, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: Targeting notch signaling to restore neural development and behavior in mouse models of ASD

doi: 10.1038/s41467-026-70321-6

Figure Lengend Snippet: a , b Flow cytometric analysis comparing Notch1 expression in the GE and the cortex ( a ), and in the transient VPA-treated forebrain primary culture (in vitro ASD model) and untreated culture ( b ). c Quantification of active NOTCH1 intracellular domain (ICN) protein levels in the in vitro ASD model, with or without pharmacological inhibition of Notch activity by DAPT treatment (Control= 7, VPA = 9, VPA + DAPT = 9, DAPT = 5). P = 0.0087 (Control vs. VPA), p < 0.0001 (VPA vs. VPA + DAPT). d Experimental scheme for genetic and pharmacological manipulation of Notch signaling in forebrain culture. Notch inhibition or activation was induced on DIV3–4, and maintained until DIV22–26. e , f Effects of Notch gain-of-function and loss-of-function on Vip transcript levels using the in vitro ASD models. e Overexpression of Notch1 ICN via retroviral infection reduced Vip transcript levels in forebrain cultures (Control = 9, ICN = 15). P = 0.0017 ( Vip ), p = 0.2867 ( Sst ). f Genome editing-mediated Notch1 gene disruption or DAPT treatment increased Vip transcript levels (Control = 5, sgN1 = 6; Control = 6, DAPT = 3). Single-guide RNAs of Notch1 (sgN1) was introduced with retrovirus vectors into forebrain cultures. The cultures used in the genome-editing experiments were prepared from embryos of Cas9-expressing mice. P = 0.0066 ( Vip , sgN1), p = 0.6345 ( Sst , sgN1), p < 0.0001 ( Vip , DAPT), p = 0.0736 ( Sst , DAPT). g Rescue of VPA-induced reduction in Vip transcript through Notch1 gene disruption (sgN1) or DAPT treatment (Control = 11, VPA = 21, VPA+sgN1 = 12; Control = 14, VPA = 15, VPA + DAPT = 4). P < 0.0001. h , i Representative images and quantification of VIP-IN density in forebrain cultures, showing significant reduction following overexpression of Notch1 ICN (Control = 9, ICN = 7). Arrowheads indicate VIP-stained cells. P = 0.0027. Scale bar: 50 µm. j , k Representative images and quantification of VIP-IN density showing restoration by gene disruption of Notch1 in VPA-exposed forebrain cultures prepared from Cas9-expressing embryos (Control = 7, VPA = 7, VPA+sgN1 = 15). P < 0.0001 (Control vs. VPA), p = 0.0003 (VPA vs. VPA+ sgN1). Scale bar: 50 µm. c , e , f , g , k , i Data are presented as mean values +/− SEM. c , g , k One-way ANOVA with Bonferroni’s multiple comparisons test. e , f , i Two-tailed unpaired t-test. Source data are provided as a Source Data file.

Article Snippet: The following commercially available primary antibodies were used: Rabbit anti-VIP (1:1000, 20077, ImmunoStar) Mouse anti-Reelin (1:500, D223-3 (CR-50), MBL) Rat anti-Somatostatin (1:300, MAB354 (YC7), Merck Millipore) Rabbit anti-Parvalbumin (1:1000, MSFR105210, Nittobo Medical) Rabbit anti-Calretinin (1:1000, MSFR100440, Nittobo Medical) Mouse anti-Calretinin (1:1000, MAB1568 (6B8.2), Merck Millipore) Mouse anti-GAD67 (1:1000, MAB5406 (1G10.2), Merck Millipore) Rabbit anti-PROX1 (1:1000, ab199359, Abcam) Mouse anti-COUP-TFII (1:1000, PP-H7147-00 (H7147), Perseus Proteomics) Rabbit anti-SOX6 (1:1000, ab30455, Abcam) Mouse anti-LHX6 (1:1000, H00026468-M02 (1B11), Abnova) Mouse anti-HDAC3 (1:1000, #3949 (7G6C5), Cell Signaling) Rabbit anti-cleaved NOTCH1 (1:1000, #4147 (Val1744) (D3D8), Cell Signaling,) Mouse anti-GAPDH (1:1000, Santa Cruz Biotechnology) For immunostaining, Alexa Fluor 488- or 546-conjugated secondary antibodies (Molecular Probes) were used.

Techniques: Expressing, In Vitro, Inhibition, Activity Assay, Control, Activation Assay, Over Expression, Retroviral, Infection, Disruption, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Targeting notch signaling to restore neural development and behavior in mouse models of ASD

doi: 10.1038/s41467-026-70321-6

Figure Lengend Snippet: a Generation of CGE-specific Notch1/2 cKO mice. Htr3a -Cre mice were used to delete Notch1/2 genes in CGE progenitors exposed to VPA. b Restoration of VIP-IN density in VPA-exposed cKO mice. Representative coronal images of the somatosensory cortex and quantification of VIP-IN density (control=46, cKO=45, WT-VPA = 45, cKO-VPA = 46 images). P < 0.0001 (WT vs. WT-VPA, cKO vs. WT-VPA, WT-VPA vs. cKO-VPA). Scale bar: 100 µm. c – g Behavioral rescue in VPA-exposed cKO mice. c Duration of self-grooming (control=30, cKO=23, WT-VPA = 44, cKO-VPA = 36). P > 0.9999 (WT vs. cKO), p = 0.0018 (WT vs. WT-VPA), p = 0.0047 (WT vs. cKO-VPA), p > 0.9999 (WT-VPA vs. cKO-VPA). d Contact time in the reciprocal social interaction test (control=29, cKO=25, WT-VPA = 32, cKO-VPA = 26). P = 0.0006 (WT vs. WT-VPA), p < 0.0001 (cKO vs. WT-VPA), p = 0.0476 (WT-VPA vs. cKO-VPA). e – g Three-chamber social interaction test (control = 57, cKO=54, WT-VPA = 63, cKO-VPA = 51). e Time spent in each chamber. Increased time in the familiar chamber by VPA-exposure, p = 0.0447 (WT vs. WT-VPA), p = 0.0128 (cKO vs. WT-VPA), p = 0.0004 (WT-VPA vs. cKO-VPA). Decreased time in the centre chamber. p = 0.0154 (cKO vs. WT-VPA), p = 0.0252 (WT-VPA vs. cKO-VPA). f Representative heatmaps of each test mouse. g SNI in the social novelty session. P = 0.0378 (WT vs. WT-VPA), p = 0.0205 (cKO vs. WT-VPA), p = 0.0021 (WT-VPA vs. cKO-VPA). b , c , d , e , g Data are presented as mean values +/−SEM. b , c , d , g One-way ANOVA with Bonferroni’s multiple comparisons test. e Two-tailed unpaired t-test. h – m Transcriptomic changes analyzed by RNA-seq of the adult forebrain ( n = 3) exhibited gene expression profiles between four groups (WT, WT-VPA, cKO, and cKO-VPA). h Number of differentially altered genes across genotypes and/or condition. i PCA showed distinct clustering of cKO-VPA mice from WT-VPA mice. j Gene enrichment analysis by STRING revealed increased glutamatergic synaptic transmission in WT-VPA, restored in cKO-VPA. k – m Volcano plots illustrating differential gene expression in comparisons: WT vs. cKO ( k ), WT vs. WT-VPA ( l ), and WT-VPA vs. cKO-VPA ( m ), highlighting the selective downregulation of Glutamatergic neuron-related genes in the cKO-VPA mice. j – m Two-sided unpaired t-test with Benjamini–Hochberg multiple testing correction. Source data are provided as a Source Data file.

Article Snippet: The following commercially available primary antibodies were used: Rabbit anti-VIP (1:1000, 20077, ImmunoStar) Mouse anti-Reelin (1:500, D223-3 (CR-50), MBL) Rat anti-Somatostatin (1:300, MAB354 (YC7), Merck Millipore) Rabbit anti-Parvalbumin (1:1000, MSFR105210, Nittobo Medical) Rabbit anti-Calretinin (1:1000, MSFR100440, Nittobo Medical) Mouse anti-Calretinin (1:1000, MAB1568 (6B8.2), Merck Millipore) Mouse anti-GAD67 (1:1000, MAB5406 (1G10.2), Merck Millipore) Rabbit anti-PROX1 (1:1000, ab199359, Abcam) Mouse anti-COUP-TFII (1:1000, PP-H7147-00 (H7147), Perseus Proteomics) Rabbit anti-SOX6 (1:1000, ab30455, Abcam) Mouse anti-LHX6 (1:1000, H00026468-M02 (1B11), Abnova) Mouse anti-HDAC3 (1:1000, #3949 (7G6C5), Cell Signaling) Rabbit anti-cleaved NOTCH1 (1:1000, #4147 (Val1744) (D3D8), Cell Signaling,) Mouse anti-GAPDH (1:1000, Santa Cruz Biotechnology) For immunostaining, Alexa Fluor 488- or 546-conjugated secondary antibodies (Molecular Probes) were used.

Techniques: Control, Two Tailed Test, RNA Sequencing, Gene Expression, Transmission Assay

Journal: Nature Communications

Article Title: Targeting notch signaling to restore neural development and behavior in mouse models of ASD

doi: 10.1038/s41467-026-70321-6

Figure Lengend Snippet: a Experimental scheme showing the in vivo rescue of Notch signaling by a single dose of γ-secretase inhibitor Ro4929097 (Ro) in the ASD-model mice exposure to VPA. b Normalized protein level of active Notch1 in the VPA-exposed mouse embryos by Ro treatment. c , d Rescue of VIP-IN density in Ro-treated VPA-exposed mice. c Representative coronal images of the somatosensory cortex. d Quantification of VIP-IN density showed a significant increase by Ro injection in VPA-exposed mice (control = 48, VPA = 68, VPA+Ro = 86 images). P < 0.0001 (Control vs. VPA, VPA vs. VPA+Ro). Scale bar: 100 µm. e – i Behavioral improvements in Ro-treated VPA-exposed mice. e Hair loss observed in 27% of VPA-exposed mice rescued by Ro exposure (control = 30, VPA = 34, VPA+Ro=29). P = 0.0024 (Control vs. VPA), p = 0.0414 (VPA vs. VPA+Ro). f Total grooming duration (control=70, VPA = 72, VPA+Ro=66). P < 0.0001 (Control vs. VPA), p = 0.0008 (VPA vs. VPA+Ro). g – i Reciprocal social interaction test (control=29, VPA = 41, VPA+Ro = 45). g Contact number, p = 0.0028 (Control vs. VPA), p = 0.0438 (VPA vs. VPA+Ro). h Contact time, p = 0.0034 (Control vs. VPA), p = 0.0375 (VPA vs. VPA+Ro). i Resting time, p < 0.0001 (Control vs. VPA, VPA vs. VPA+Ro). d , f , g – i Data are presented as mean values +/−SEM. One-way ANOVA with Bonferroni’s multiple comparisons test. e Two-sided Pearson’s chi-square test. j – o Single-cell RNA sequencing (scRNA-seq) analysis of whole brain (P2) from VPA-exposed mice revealed cellular and transcriptomic rescue by Ro. j UMAP plot of 20 clusters in whole-brain cells. k Restored perturbations in the whole cell compositions in VPA-exposed mouse with Ro exposure. l Heatmap of residuals and Pearson’s coefficients for cell-type distributions. Combined heatmap showing standardized residuals from chi-square tests and Pearson’s coefficients between groups (control, VPA, VPA+Ro). Ro treatment shifted the VPA-induced distribution pattern toward that of control mouse. m Sub-clustering of NSC/NP-enriched populations (clusters #6 and #7) and the normalized cellular composition of Htr3a + CGE-enriched cell populations in VPA-exposed mouse after Ro treatment. n Normalized cell composition of oligodendrocyte-enriched clusters (clusters #5, #8, #12, #14, and #18) with Ro exposure. ovgy Cell composition of astrocyte-enriched clusters (clusters #2, #3, and #13) with Ro exposure. Source data are provided as a Source Data file.

Article Snippet: The following commercially available primary antibodies were used: Rabbit anti-VIP (1:1000, 20077, ImmunoStar) Mouse anti-Reelin (1:500, D223-3 (CR-50), MBL) Rat anti-Somatostatin (1:300, MAB354 (YC7), Merck Millipore) Rabbit anti-Parvalbumin (1:1000, MSFR105210, Nittobo Medical) Rabbit anti-Calretinin (1:1000, MSFR100440, Nittobo Medical) Mouse anti-Calretinin (1:1000, MAB1568 (6B8.2), Merck Millipore) Mouse anti-GAD67 (1:1000, MAB5406 (1G10.2), Merck Millipore) Rabbit anti-PROX1 (1:1000, ab199359, Abcam) Mouse anti-COUP-TFII (1:1000, PP-H7147-00 (H7147), Perseus Proteomics) Rabbit anti-SOX6 (1:1000, ab30455, Abcam) Mouse anti-LHX6 (1:1000, H00026468-M02 (1B11), Abnova) Mouse anti-HDAC3 (1:1000, #3949 (7G6C5), Cell Signaling) Rabbit anti-cleaved NOTCH1 (1:1000, #4147 (Val1744) (D3D8), Cell Signaling,) Mouse anti-GAPDH (1:1000, Santa Cruz Biotechnology) For immunostaining, Alexa Fluor 488- or 546-conjugated secondary antibodies (Molecular Probes) were used.

Techniques: In Vivo, Injection, Control, Single Cell, RNA Sequencing

Journal: JID Innovations

Article Title: Spatiotemporal fluorescence imaging of microRNA activity in 3-D models of human epidermis reveals contribution of the Notch pathway in the regulation of miR-30a in aging skin

doi: 10.1016/j.xjidi.2025.100444

Figure Lengend Snippet: The inhibition of the Notch pathway increases miR-30a activity. (a ) Representative images of reconstructed epidermises obtained using RIFES miR-30a-3p or miR-30a-5p primary keratinocytes. The green fluorescence corresponds to GFP, and the blue fluorescence corresponds to nuclei labeled with DAPI. The limit of the epidermis is indicated by a dotted line. Bar = 50 mm. ( b ) Immunofluorescence labeling of Notch 1 (denoted as NICD) in a skin biopsy (female abdominal skin, young adult). The red fluorescence corresponds to Notch1, the green fluorescence corresponds to loricrin, and the blue fluorescence corresponds to nuclei labeled with DAPI. Bar = 50 mm. ( c ) Western blot analysis of Notch1 (denoted as NICD), cleaved Notch1, and actin in HPKs treated or not by DAPT. ( d ) HEY1 transcript relative expression analysis by qRT-PCR in HPKs treated or not by DAPT (mean ± SD, n = 3, ∗ P < .05 t -test P -value). ( e ) Response of the RIFES miR-30a-3p or -5p HPKs to DAPT treatment. The graph corresponds to the quantification of the GFP fluorescence by HCS after DAPT or control treatment (box plot with Tukey whiskers, n = 4, ∗ P < .05 t -test P -value). Representative images of cells are shown (Merge image: GFP plus DAPI). ( f ) Western blot analysis of GFP and actin expression in lentiRIFES/miR-30a-3pT or 5pT HPKs after DAPT treatment. ( g ) Reconstructed epidermises were obtained using lentiRIFES/miR-30a-3pT or 5pT keratinocytes. The RHEs were treated or not by the DAPT compound. Representative images of GFP fluorescence or K10 immunofluorescence are shown. The limit of the epidermis is indicated by a dotted line. Bar = 50 mm. A small area of the images surrounded by a frame is shown at higher magnification. ( h ) miR-30a-3p or miR-30a-5p transcript expression analysis by qRT-PCR in HPKs treated or not by DAPT (mean ± SD, n = 3, ∗ P < .05 t -test P -value). HCS, high-content screening; HPK, human primary keratinocyte; K10, keratin 10; RHE, reconstructed human epidermis.

Article Snippet: Membranes were blocked with 10% nonfat dry milk for 1 hour and incubated with primary antibodies against NOTCH 1 (number 3608, Cell Signaling Technology), cleaved NOTCH-1 (number 4147, Cell Signaling Technology), c-MYC, or β-actin (ab8227, Abcam) overnight at 4 °C.

Techniques: Inhibition, Activity Assay, Fluorescence, Labeling, Immunofluorescence, Western Blot, Expressing, Quantitative RT-PCR, Control, High Content Screening

Journal: Scientific Reports

Article Title: Caveolin-1 modulates Notch transcriptional activity during in vitro respiratory multiciliated cell maturation

doi: 10.1038/s41598-026-40201-6

Figure Lengend Snippet: Analysis of Notch1 Expression and Subcellular Distribution in Luc-KD and Cav1-KD Cells. ( a ) Maximal projection of confocal images for Notch1 (in yellow), and nucleus (in blue) in Luc-KD and Cav1-KD cells in ALI 6. ( b ) Western blot images of Luc-KD and Cav1-KD cells showing Notch 1 (full-length at 300 kDa and cleaved at 120 kDa) and E-Cadherin expression. The lower Western blot image of Notch1 (300 kDa) in the panel shows a contrast-enhanced version of Notch1 region of interest (ROI) amplified to the entire image, derived from the original image with lower contrast shown in the upper part of the panel. Each condition was tested in triplicate (L1, L2, and L3 for each genotype). Cell lysates (L) were derived from an independent cell culture. β-actin was used as a loading control. ( c – e ) Relative protein expression levels quantification of Notch1 300 kDa ( c ), Notch1 120 kDa ( d ) and E-Cadherin ( e ). Mean and standard deviation as error bars were plotted, n = 3 independent lysates per group. ( f ) Analyses of Notch 1 full-length and processed forms (TMD+NICD and NICD) subcellular distribution in the cytosol (Cyt.), membrane (Mem.), and chromatin (Chr.) in Luc-KD and Cav1-KD cells. Cell fractionation and gradient SDS-Gels were used for improved resolution. ( g ) Relative protein expression quantification of Notch 1 subcellular distribution in Luc-KD and Cav1-KD cells. Mean and standard deviation as error bars were plotted, n = 4 independent lysates per group. ( h ) Proposed working model of the mechanism by which Cav-1 regulates BSC differentiation, involving differential NICD binding capacity to chromatin together with other partners. Scale bar in panel a represent 20 μm. p-values in all conditions were obtained using two-tailed t-test (*** represents p < 0.001, ** represents p < 0.01 and n.s. means no significative differences).

Article Snippet: The remaining chromatin was diluted, precleared with protein A/G Sepharose beads, and immunoprecipitated overnight at 4 °C using an anti-NICD antibody (Cell Signaling, #4147S, 2.5 μL of antibody/10 μg of chromatin).

Techniques: Expressing, Western Blot, Amplification, Derivative Assay, Cell Culture, Control, Standard Deviation, Membrane, Cell Fractionation, Binding Assay, Two Tailed Test

Journal: Scientific Reports

Article Title: Caveolin-1 modulates Notch transcriptional activity during in vitro respiratory multiciliated cell maturation

doi: 10.1038/s41598-026-40201-6

Figure Lengend Snippet: Analysis of Notch2 Expression and Subcellular Distribution in Luc-KD and Cav1-KD Cells. ( a ) Maximal projection of confocal images for Notch2 (in green), and nucleus (in blue) in Luc-KD and Cav1-KD cells in ALI 6. ( b ) Western blot images of Luc-KD and Cav1-KD cells showing Notch 2 (full-length at 300 kDa and cleaved at 110 kDa) and c-Myb expression. Each condition was tested in triplicate (L1, L2, and L3 for each genotype). Cell lysates (L) were derived from an independent cell culture. GAPDH was used as a loading control. ( c – e ) Relative protein expression levels quantification of Notch2 300 kDa ( c ), Notch2 120 kDa ( d ) and c-Myb ( e ). Mean and standard deviation as error bars were plotted, n = 3 independent lysates per group. ( f ) Analyses of Notch 2 full-length and processed forms (TMD+NICD and NICD) subcellular distribution in the cytosol (Cyt.), membrane (Mem.), and chromatin (Chr.) in Luc-KD and Cav1-KD cells. Cell fractionation and gradient SDS-Gels were used for improved resolution. ( g ) Relative protein expression quantification of Notch 2 subcellular distribution in Luc-KD and Cav1-KD cells. Mean and standard deviation as error bars were plotted, n = 4 independent lysates per group. Scale bar in panel a represent 20 μm. p-values in all conditions were obtained using two-tailed t-test (**represents p < 0.01, *represents p < 0.05 and n.s. means no significative differences).

Article Snippet: The remaining chromatin was diluted, precleared with protein A/G Sepharose beads, and immunoprecipitated overnight at 4 °C using an anti-NICD antibody (Cell Signaling, #4147S, 2.5 μL of antibody/10 μg of chromatin).

Techniques: Expressing, Western Blot, Derivative Assay, Cell Culture, Control, Standard Deviation, Membrane, Cell Fractionation, Two Tailed Test