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human notch1  (MedChemExpress)


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    Structured Review

    MedChemExpress human notch1
    (A-D) Box and scatter plots showing bulk RNA-sequencing data for key Notch pathway genes across a panel of human prostate cancer cell lines (LNCaP, LNCaP-abl, C4-2, C4-2B, VCaP, 22RV1, PC3, DU145, H660, LASCPC1, LNCaP_42D). (A-B) <t>NOTCH1</t> and NOTCH2 Gene Expression (receptor). (C) HES1 Gene Expression (downstream effector). (D) DLL3 Gene Expression (ligand). (E) Western blot analysis confirming the protein expression of NOTCH2, DLL3, and HES1 in selected cell lines (LNCaP/AR, PC3, gTP53/RB1-R, and LASCPC1), corroborating the gene expression trends observed in the bulk RNA-seq data. Actin serves as a loading control.
    Human Notch1, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human notch1/product/MedChemExpress
    Average 94 stars, based on 24 article reviews
    human notch1 - by Bioz Stars, 2026-02
    94/100 stars

    Images

    1) Product Images from "Graded Notch Signaling Functions as a Rheostat of Lineage Plasticity and Therapy Resistance in Prostate Cancer"

    Article Title: Graded Notch Signaling Functions as a Rheostat of Lineage Plasticity and Therapy Resistance in Prostate Cancer

    Journal: bioRxiv

    doi: 10.1101/2025.11.23.690056

    (A-D) Box and scatter plots showing bulk RNA-sequencing data for key Notch pathway genes across a panel of human prostate cancer cell lines (LNCaP, LNCaP-abl, C4-2, C4-2B, VCaP, 22RV1, PC3, DU145, H660, LASCPC1, LNCaP_42D). (A-B) NOTCH1 and NOTCH2 Gene Expression (receptor). (C) HES1 Gene Expression (downstream effector). (D) DLL3 Gene Expression (ligand). (E) Western blot analysis confirming the protein expression of NOTCH2, DLL3, and HES1 in selected cell lines (LNCaP/AR, PC3, gTP53/RB1-R, and LASCPC1), corroborating the gene expression trends observed in the bulk RNA-seq data. Actin serves as a loading control.
    Figure Legend Snippet: (A-D) Box and scatter plots showing bulk RNA-sequencing data for key Notch pathway genes across a panel of human prostate cancer cell lines (LNCaP, LNCaP-abl, C4-2, C4-2B, VCaP, 22RV1, PC3, DU145, H660, LASCPC1, LNCaP_42D). (A-B) NOTCH1 and NOTCH2 Gene Expression (receptor). (C) HES1 Gene Expression (downstream effector). (D) DLL3 Gene Expression (ligand). (E) Western blot analysis confirming the protein expression of NOTCH2, DLL3, and HES1 in selected cell lines (LNCaP/AR, PC3, gTP53/RB1-R, and LASCPC1), corroborating the gene expression trends observed in the bulk RNA-seq data. Actin serves as a loading control.

    Techniques Used: RNA Sequencing, Gene Expression, Western Blot, Expressing, Control

    (A, B) Relative gene expression of Notch pathway components and lineage markers in enzalutamide-resistant cell lines, (A) gTP53/RB1-R and (B) PC3, after siRNA knockdown of specific genes ( NOTCH1, NOTCH2, HES1 ) compared to non-targeting control (NC). Significance is denoted as *p < 0.05, **p < 0.01, ***p < 0.001.
    Figure Legend Snippet: (A, B) Relative gene expression of Notch pathway components and lineage markers in enzalutamide-resistant cell lines, (A) gTP53/RB1-R and (B) PC3, after siRNA knockdown of specific genes ( NOTCH1, NOTCH2, HES1 ) compared to non-targeting control (NC). Significance is denoted as *p < 0.05, **p < 0.01, ***p < 0.001.

    Techniques Used: Gene Expression, Knockdown, Control



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    (A-D) Box and scatter plots showing bulk RNA-sequencing data for key Notch pathway genes across a panel of human prostate cancer cell lines (LNCaP, LNCaP-abl, C4-2, C4-2B, VCaP, 22RV1, PC3, DU145, H660, LASCPC1, LNCaP_42D). (A-B) <t>NOTCH1</t> and NOTCH2 Gene Expression (receptor). (C) HES1 Gene Expression (downstream effector). (D) DLL3 Gene Expression (ligand). (E) Western blot analysis confirming the protein expression of NOTCH2, DLL3, and HES1 in selected cell lines (LNCaP/AR, PC3, gTP53/RB1-R, and LASCPC1), corroborating the gene expression trends observed in the bulk RNA-seq data. Actin serves as a loading control.
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    (A-D) Box and scatter plots showing bulk RNA-sequencing data for key Notch pathway genes across a panel of human prostate cancer cell lines (LNCaP, LNCaP-abl, C4-2, C4-2B, VCaP, 22RV1, PC3, DU145, H660, LASCPC1, LNCaP_42D). (A-B) <t>NOTCH1</t> and NOTCH2 Gene Expression (receptor). (C) HES1 Gene Expression (downstream effector). (D) DLL3 Gene Expression (ligand). (E) Western blot analysis confirming the protein expression of NOTCH2, DLL3, and HES1 in selected cell lines (LNCaP/AR, PC3, gTP53/RB1-R, and LASCPC1), corroborating the gene expression trends observed in the bulk RNA-seq data. Actin serves as a loading control.
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    AKT1 Enhances <t>Notch1</t> Nuclear Translocation via Phosphorylation. A : Co-IP confirms the interaction between Notch1 and AKT1. B : In vitro kinase assays reveal AKT1-induced Notch1 phosphorylation. C : A stable MGC803 cell line overexpressing AKT1 is treated ± LY294002; Western Blot assesses Flag-tagged AKT1 levels and IP measures Notch1’s RXX(S*/T*) motif phosphorylation. D : Statistical analysis of phospho-Akt substrate expression in Notch1 from C. E - H : After AKT1 overexpression and LY294002 treatment, subcellular fractionation is performed, and Western Blot analyzes Notch1 and Notch1-IC levels, with subsequent statistical analysis. I : Phosphoprotemic analysis identifies the phosphorylation site on Notch1. J : Immunofluorescence tests AKT1’s effect on Notch1 nuclear localization (×200). K : The quantification of Notch signal intensity in the nucleus. n = 3. Significance: vs. NC group, # P < 0.05, ## P < 0.01, ### P < 0.001; vs. oe-Myr-AKT1 group, * P < 0.05, *** P < 0.001
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    Image Search Results


    (A-D) Box and scatter plots showing bulk RNA-sequencing data for key Notch pathway genes across a panel of human prostate cancer cell lines (LNCaP, LNCaP-abl, C4-2, C4-2B, VCaP, 22RV1, PC3, DU145, H660, LASCPC1, LNCaP_42D). (A-B) NOTCH1 and NOTCH2 Gene Expression (receptor). (C) HES1 Gene Expression (downstream effector). (D) DLL3 Gene Expression (ligand). (E) Western blot analysis confirming the protein expression of NOTCH2, DLL3, and HES1 in selected cell lines (LNCaP/AR, PC3, gTP53/RB1-R, and LASCPC1), corroborating the gene expression trends observed in the bulk RNA-seq data. Actin serves as a loading control.

    Journal: bioRxiv

    Article Title: Graded Notch Signaling Functions as a Rheostat of Lineage Plasticity and Therapy Resistance in Prostate Cancer

    doi: 10.1101/2025.11.23.690056

    Figure Lengend Snippet: (A-D) Box and scatter plots showing bulk RNA-sequencing data for key Notch pathway genes across a panel of human prostate cancer cell lines (LNCaP, LNCaP-abl, C4-2, C4-2B, VCaP, 22RV1, PC3, DU145, H660, LASCPC1, LNCaP_42D). (A-B) NOTCH1 and NOTCH2 Gene Expression (receptor). (C) HES1 Gene Expression (downstream effector). (D) DLL3 Gene Expression (ligand). (E) Western blot analysis confirming the protein expression of NOTCH2, DLL3, and HES1 in selected cell lines (LNCaP/AR, PC3, gTP53/RB1-R, and LASCPC1), corroborating the gene expression trends observed in the bulk RNA-seq data. Actin serves as a loading control.

    Article Snippet: A pool of siRNAs targeting human NOTCH1, NOTCH2, HES1, and a non-targeting control pool were used (MedChemExpress; HY-RS09445, HY-RS09448, HY-RS06134).

    Techniques: RNA Sequencing, Gene Expression, Western Blot, Expressing, Control

    (A, B) Relative gene expression of Notch pathway components and lineage markers in enzalutamide-resistant cell lines, (A) gTP53/RB1-R and (B) PC3, after siRNA knockdown of specific genes ( NOTCH1, NOTCH2, HES1 ) compared to non-targeting control (NC). Significance is denoted as *p < 0.05, **p < 0.01, ***p < 0.001.

    Journal: bioRxiv

    Article Title: Graded Notch Signaling Functions as a Rheostat of Lineage Plasticity and Therapy Resistance in Prostate Cancer

    doi: 10.1101/2025.11.23.690056

    Figure Lengend Snippet: (A, B) Relative gene expression of Notch pathway components and lineage markers in enzalutamide-resistant cell lines, (A) gTP53/RB1-R and (B) PC3, after siRNA knockdown of specific genes ( NOTCH1, NOTCH2, HES1 ) compared to non-targeting control (NC). Significance is denoted as *p < 0.05, **p < 0.01, ***p < 0.001.

    Article Snippet: A pool of siRNAs targeting human NOTCH1, NOTCH2, HES1, and a non-targeting control pool were used (MedChemExpress; HY-RS09445, HY-RS09448, HY-RS06134).

    Techniques: Gene Expression, Knockdown, Control

    Fig. 2 siRNA-PTX-NPs suppressed NOTCH1 and SF3B1 at both the mRNA and protein levels. NOTCH1 and SF3B1 mRNA and protein expression levels were measured using qRT-PCR (a, b) and colorimetric analysis after treatment with various therapeutic agents (c-d). The data were normalized using β-actin mRNA expression levels as an internal control. The 2 –ΔΔCt technique was exploited to compute the relative mRNA expression level of the targeted genes. P-values < 0.05 (*), P-values < 0.01 (**), and P-values < 0.001 (***)

    Journal: Cancer cell international

    Article Title: Advancing the therapeutic effectiveness of paclitaxel in chronic lymphocytic leukemia through the simultaneous inhibition of NOTCH1 and SF3B1.

    doi: 10.1186/s12935-025-03702-4

    Figure Lengend Snippet: Fig. 2 siRNA-PTX-NPs suppressed NOTCH1 and SF3B1 at both the mRNA and protein levels. NOTCH1 and SF3B1 mRNA and protein expression levels were measured using qRT-PCR (a, b) and colorimetric analysis after treatment with various therapeutic agents (c-d). The data were normalized using β-actin mRNA expression levels as an internal control. The 2 –ΔΔCt technique was exploited to compute the relative mRNA expression level of the targeted genes. P-values < 0.05 (*), P-values < 0.01 (**), and P-values < 0.001 (***)

    Article Snippet: Specific human siRNA against NOTCH1, SF3B1, and control siRNA were acquired from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Expressing, Quantitative RT-PCR, Control

    Fig. 4 Silencing of SF3B1 and NOTCH1 significantly enhances the sensitivity of CLL cells to PTX-induced apoptosis. ELISA method was used to determine the cell death after treatment of CLL cells with different therapeutic agents (a). The qRT-PCR technique measured the mRNA expression levels of Bcl-2 and Bax after treatment of both PBMCs and BMMCs with different therapeutic agents (b, c). P-values < 0.05 (*), P-values < 0.01 (**), and P-values < 0.001 (***)

    Journal: Cancer cell international

    Article Title: Advancing the therapeutic effectiveness of paclitaxel in chronic lymphocytic leukemia through the simultaneous inhibition of NOTCH1 and SF3B1.

    doi: 10.1186/s12935-025-03702-4

    Figure Lengend Snippet: Fig. 4 Silencing of SF3B1 and NOTCH1 significantly enhances the sensitivity of CLL cells to PTX-induced apoptosis. ELISA method was used to determine the cell death after treatment of CLL cells with different therapeutic agents (a). The qRT-PCR technique measured the mRNA expression levels of Bcl-2 and Bax after treatment of both PBMCs and BMMCs with different therapeutic agents (b, c). P-values < 0.05 (*), P-values < 0.01 (**), and P-values < 0.001 (***)

    Article Snippet: Specific human siRNA against NOTCH1, SF3B1, and control siRNA were acquired from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing

    Fig. 6 The competing endogenous RNA (ceRNA) network that modulates NOTCH1 and SF3B1 genes. The ceRNA network, including NOTCH1 and SF3B1, 9 miRNAs targeting both genes and 46 lncRNAs sponging selected miRNAs were constructed (a). The enrichment analysis underscored the pivotal path ways where the ceRNA network, associated with NOTCH1 and SF3B1, exerts its influence in the context of chronic lymphocytic leukemia (b)

    Journal: Cancer cell international

    Article Title: Advancing the therapeutic effectiveness of paclitaxel in chronic lymphocytic leukemia through the simultaneous inhibition of NOTCH1 and SF3B1.

    doi: 10.1186/s12935-025-03702-4

    Figure Lengend Snippet: Fig. 6 The competing endogenous RNA (ceRNA) network that modulates NOTCH1 and SF3B1 genes. The ceRNA network, including NOTCH1 and SF3B1, 9 miRNAs targeting both genes and 46 lncRNAs sponging selected miRNAs were constructed (a). The enrichment analysis underscored the pivotal path ways where the ceRNA network, associated with NOTCH1 and SF3B1, exerts its influence in the context of chronic lymphocytic leukemia (b)

    Article Snippet: Specific human siRNA against NOTCH1, SF3B1, and control siRNA were acquired from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Construct

    AKT1 Enhances Notch1 Nuclear Translocation via Phosphorylation. A : Co-IP confirms the interaction between Notch1 and AKT1. B : In vitro kinase assays reveal AKT1-induced Notch1 phosphorylation. C : A stable MGC803 cell line overexpressing AKT1 is treated ± LY294002; Western Blot assesses Flag-tagged AKT1 levels and IP measures Notch1’s RXX(S*/T*) motif phosphorylation. D : Statistical analysis of phospho-Akt substrate expression in Notch1 from C. E - H : After AKT1 overexpression and LY294002 treatment, subcellular fractionation is performed, and Western Blot analyzes Notch1 and Notch1-IC levels, with subsequent statistical analysis. I : Phosphoprotemic analysis identifies the phosphorylation site on Notch1. J : Immunofluorescence tests AKT1’s effect on Notch1 nuclear localization (×200). K : The quantification of Notch signal intensity in the nucleus. n = 3. Significance: vs. NC group, # P < 0.05, ## P < 0.01, ### P < 0.001; vs. oe-Myr-AKT1 group, * P < 0.05, *** P < 0.001

    Journal: Journal of Cancer Research and Clinical Oncology

    Article Title: AKT1-Mediated NOTCH1 phosphorylation promotes gastric cancer progression via targeted regulation of IRS-1 transcription

    doi: 10.1007/s00432-024-06039-z

    Figure Lengend Snippet: AKT1 Enhances Notch1 Nuclear Translocation via Phosphorylation. A : Co-IP confirms the interaction between Notch1 and AKT1. B : In vitro kinase assays reveal AKT1-induced Notch1 phosphorylation. C : A stable MGC803 cell line overexpressing AKT1 is treated ± LY294002; Western Blot assesses Flag-tagged AKT1 levels and IP measures Notch1’s RXX(S*/T*) motif phosphorylation. D : Statistical analysis of phospho-Akt substrate expression in Notch1 from C. E - H : After AKT1 overexpression and LY294002 treatment, subcellular fractionation is performed, and Western Blot analyzes Notch1 and Notch1-IC levels, with subsequent statistical analysis. I : Phosphoprotemic analysis identifies the phosphorylation site on Notch1. J : Immunofluorescence tests AKT1’s effect on Notch1 nuclear localization (×200). K : The quantification of Notch signal intensity in the nucleus. n = 3. Significance: vs. NC group, # P < 0.05, ## P < 0.01, ### P < 0.001; vs. oe-Myr-AKT1 group, * P < 0.05, *** P < 0.001

    Article Snippet: Fetal bovine serum (FBS, Beijing Cellmax Company, SA111.02); RPMI-1640 (Gibco, 42401042); DMEM (Dalian Meilun Biology, MA0212-2); LipofiterTM Transfection Reagent (Hanheng, HB-TRCF-1000); RIPA Lysis buffer (Beijing Solarbio Company, R0010); SDS Lysis Buffer (Beyotime, P0013G); Membrane Protein Extraction Kit (Solarbio, EX1500); Nuclear Protein Extraction Kit (Solarbio, EX1470); Notch1 Antibody (CST, 3447); Recombinant AKT1 Protein (Abcam, ab79792); Notch1-IC Antibody (CST, 4147); AKT1 Antibody (Abcam, ab238477); phospho-Akt substrate RXX(S*/T*) Antibody (CST, 9614); IRS-1 Antibody (Abcam, ab131487); Flag Antibody (Abcam, ab205606); E-cadherin Antibody(Abcam, ab231303); Snail1 Atibody(CST, 3879); Vimentin Antibody(Abcam, ab92547); Co-IP Kit (Thermo, 26149); Kinase Buffer (CST, 9802); ATP (CST, 9804); Recombinant human AKT1 protein (Active) (Abcam, ab62279); Recombinant Human NOTCH1 protein (hFc Tag) (Sinobiological, 10954-H02H); TRIzol (Ambion, USA, 15596026); PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) (TaKaRa, Japan, RR047A); TB Green ® Premix Ex Taq™ II (Tli RNaseH Plus) (TaKaRa, Japan, RR820A); PMSF (Beijing Solarbio Company, P0100); SDS-PAGE Gel Preparation Kit (Beijing Solarbio Company, P1200); ECL Ultra-Sensitive Luminol Reagent (Beijing Solarbio Company, PE0010); Pierce Co-IP Kit(Thermo Scientific, 26149), Protein A Agarose/Salmon Sperm DNA (Sigma-Aldrich, 16–157).

    Techniques: Translocation Assay, Co-Immunoprecipitation Assay, In Vitro, Western Blot, Expressing, Over Expression, Fractionation, Immunofluorescence

    Elevated IRS-1 in Gastric Cancer Impairs Prognosis. A : Databases TRANSFAC and CHEA identified Notch1-RBPJ target genes intersecting with differentially expressed genes in gastric cancer. B : MGC803 cell line overexpressing Notch1-IC was used to assess the effect on IRS-1, CDH5, TNL1, ASCL2, and LRP6 expression via RT-PCR ( n = 3 per group), with significance marked as * P < 0.05 to **** P < 0.0001 vs. oe-NC. C : GEPIA compared IRS-1 expression in gastric cancer vs. normal tissues. D - F : IHC and RT-PCR analyzed IRS-1 expression in clinical samples, with a chi-square test for microarray positive rates ( n = 97 per group). G : TCGA data correlated IRS-1 expression with cancer stage. H : Survival analysis was performed on tissue microarray IRS-1 data

    Journal: Journal of Cancer Research and Clinical Oncology

    Article Title: AKT1-Mediated NOTCH1 phosphorylation promotes gastric cancer progression via targeted regulation of IRS-1 transcription

    doi: 10.1007/s00432-024-06039-z

    Figure Lengend Snippet: Elevated IRS-1 in Gastric Cancer Impairs Prognosis. A : Databases TRANSFAC and CHEA identified Notch1-RBPJ target genes intersecting with differentially expressed genes in gastric cancer. B : MGC803 cell line overexpressing Notch1-IC was used to assess the effect on IRS-1, CDH5, TNL1, ASCL2, and LRP6 expression via RT-PCR ( n = 3 per group), with significance marked as * P < 0.05 to **** P < 0.0001 vs. oe-NC. C : GEPIA compared IRS-1 expression in gastric cancer vs. normal tissues. D - F : IHC and RT-PCR analyzed IRS-1 expression in clinical samples, with a chi-square test for microarray positive rates ( n = 97 per group). G : TCGA data correlated IRS-1 expression with cancer stage. H : Survival analysis was performed on tissue microarray IRS-1 data

    Article Snippet: Fetal bovine serum (FBS, Beijing Cellmax Company, SA111.02); RPMI-1640 (Gibco, 42401042); DMEM (Dalian Meilun Biology, MA0212-2); LipofiterTM Transfection Reagent (Hanheng, HB-TRCF-1000); RIPA Lysis buffer (Beijing Solarbio Company, R0010); SDS Lysis Buffer (Beyotime, P0013G); Membrane Protein Extraction Kit (Solarbio, EX1500); Nuclear Protein Extraction Kit (Solarbio, EX1470); Notch1 Antibody (CST, 3447); Recombinant AKT1 Protein (Abcam, ab79792); Notch1-IC Antibody (CST, 4147); AKT1 Antibody (Abcam, ab238477); phospho-Akt substrate RXX(S*/T*) Antibody (CST, 9614); IRS-1 Antibody (Abcam, ab131487); Flag Antibody (Abcam, ab205606); E-cadherin Antibody(Abcam, ab231303); Snail1 Atibody(CST, 3879); Vimentin Antibody(Abcam, ab92547); Co-IP Kit (Thermo, 26149); Kinase Buffer (CST, 9802); ATP (CST, 9804); Recombinant human AKT1 protein (Active) (Abcam, ab62279); Recombinant Human NOTCH1 protein (hFc Tag) (Sinobiological, 10954-H02H); TRIzol (Ambion, USA, 15596026); PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) (TaKaRa, Japan, RR047A); TB Green ® Premix Ex Taq™ II (Tli RNaseH Plus) (TaKaRa, Japan, RR820A); PMSF (Beijing Solarbio Company, P0100); SDS-PAGE Gel Preparation Kit (Beijing Solarbio Company, P1200); ECL Ultra-Sensitive Luminol Reagent (Beijing Solarbio Company, PE0010); Pierce Co-IP Kit(Thermo Scientific, 26149), Protein A Agarose/Salmon Sperm DNA (Sigma-Aldrich, 16–157).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Microarray

    Activated Notch1-IC Regulates IRS-1 to Modulate Cellular Functions. A - F : Overexpression of Notch1-IC in MGC803 and HCG27 cells confirmed regulatory effects on IRS-1 by Western Blot and RT-PCR ( n = 3 per group), with significance as ** P < 0.01, *** P < 0.001 vs. oeNC. G : JASPAR predicted RBPJ binding sites in the IRS-1 promoter region. H , I : ChIP-PCR assessed the transcriptional interaction between Notch1-IC and IRS-1 in MGC803, quantifying promoter binding ( n = 3 per group), with * P < 0.05 vs. oeNC. J , K : Western Blot validated shIRS-1 lentivirus-mediated IRS-1 knockdown and performed semi-quantitative analysis ( n = 3 per group), with ** P < 0.01, *** P < 0.001 vs. shNC. L - U : Stable cell lines were created with Notch1-IC overexpression alone or with shIRS-1. EdU (L-O, ×100), Transwell (P-I, ×100), and scratch assays (S-U, ×40) measured cellular functions ( n = 3 per group), with ## P < 0.01, ### P < 0.001 vs. oeNC; * P < 0.05, ** P < 0.01, *** P < 0.001 vs. oeNotch1-IC

    Journal: Journal of Cancer Research and Clinical Oncology

    Article Title: AKT1-Mediated NOTCH1 phosphorylation promotes gastric cancer progression via targeted regulation of IRS-1 transcription

    doi: 10.1007/s00432-024-06039-z

    Figure Lengend Snippet: Activated Notch1-IC Regulates IRS-1 to Modulate Cellular Functions. A - F : Overexpression of Notch1-IC in MGC803 and HCG27 cells confirmed regulatory effects on IRS-1 by Western Blot and RT-PCR ( n = 3 per group), with significance as ** P < 0.01, *** P < 0.001 vs. oeNC. G : JASPAR predicted RBPJ binding sites in the IRS-1 promoter region. H , I : ChIP-PCR assessed the transcriptional interaction between Notch1-IC and IRS-1 in MGC803, quantifying promoter binding ( n = 3 per group), with * P < 0.05 vs. oeNC. J , K : Western Blot validated shIRS-1 lentivirus-mediated IRS-1 knockdown and performed semi-quantitative analysis ( n = 3 per group), with ** P < 0.01, *** P < 0.001 vs. shNC. L - U : Stable cell lines were created with Notch1-IC overexpression alone or with shIRS-1. EdU (L-O, ×100), Transwell (P-I, ×100), and scratch assays (S-U, ×40) measured cellular functions ( n = 3 per group), with ## P < 0.01, ### P < 0.001 vs. oeNC; * P < 0.05, ** P < 0.01, *** P < 0.001 vs. oeNotch1-IC

    Article Snippet: Fetal bovine serum (FBS, Beijing Cellmax Company, SA111.02); RPMI-1640 (Gibco, 42401042); DMEM (Dalian Meilun Biology, MA0212-2); LipofiterTM Transfection Reagent (Hanheng, HB-TRCF-1000); RIPA Lysis buffer (Beijing Solarbio Company, R0010); SDS Lysis Buffer (Beyotime, P0013G); Membrane Protein Extraction Kit (Solarbio, EX1500); Nuclear Protein Extraction Kit (Solarbio, EX1470); Notch1 Antibody (CST, 3447); Recombinant AKT1 Protein (Abcam, ab79792); Notch1-IC Antibody (CST, 4147); AKT1 Antibody (Abcam, ab238477); phospho-Akt substrate RXX(S*/T*) Antibody (CST, 9614); IRS-1 Antibody (Abcam, ab131487); Flag Antibody (Abcam, ab205606); E-cadherin Antibody(Abcam, ab231303); Snail1 Atibody(CST, 3879); Vimentin Antibody(Abcam, ab92547); Co-IP Kit (Thermo, 26149); Kinase Buffer (CST, 9802); ATP (CST, 9804); Recombinant human AKT1 protein (Active) (Abcam, ab62279); Recombinant Human NOTCH1 protein (hFc Tag) (Sinobiological, 10954-H02H); TRIzol (Ambion, USA, 15596026); PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) (TaKaRa, Japan, RR047A); TB Green ® Premix Ex Taq™ II (Tli RNaseH Plus) (TaKaRa, Japan, RR820A); PMSF (Beijing Solarbio Company, P0100); SDS-PAGE Gel Preparation Kit (Beijing Solarbio Company, P1200); ECL Ultra-Sensitive Luminol Reagent (Beijing Solarbio Company, PE0010); Pierce Co-IP Kit(Thermo Scientific, 26149), Protein A Agarose/Salmon Sperm DNA (Sigma-Aldrich, 16–157).

    Techniques: Over Expression, Western Blot, Reverse Transcription Polymerase Chain Reaction, Binding Assay, Knockdown, Stable Transfection

    AKT1-Phosphorylated Notch1 Enhances Gastric Cancer Progression via through the Regulation of IRS-1. A , B : MGC803 cells overexpressing AKT were treated with LY294002, and Western Blot confirmed AKT1’s regulation of IRS-1 expression, with semi-quantitative analysis ( n = 3 per group). Significance: ### P < 0.001 vs. oeNC; ** P < 0.01 vs. oe-Myr-AKT1. C - K : In HGC-27 and MGC803 cells with AKT1 overexpression, LY294002 and shIRS-1 lentivirus were applied to assess the effects on cell proliferation (EdU) (C-E, ×100), invasion (Transwell) ( F , H , I , ×200), and migration (scratch) (G, J, K, ×40) ( n = 3 per group). Significance: ## P < 0.01, ### P < 0.001 vs. oeNC; * P < 0.05, ** P < 0.01, *** P < 0.001 vs. oe-Myr-AKT1

    Journal: Journal of Cancer Research and Clinical Oncology

    Article Title: AKT1-Mediated NOTCH1 phosphorylation promotes gastric cancer progression via targeted regulation of IRS-1 transcription

    doi: 10.1007/s00432-024-06039-z

    Figure Lengend Snippet: AKT1-Phosphorylated Notch1 Enhances Gastric Cancer Progression via through the Regulation of IRS-1. A , B : MGC803 cells overexpressing AKT were treated with LY294002, and Western Blot confirmed AKT1’s regulation of IRS-1 expression, with semi-quantitative analysis ( n = 3 per group). Significance: ### P < 0.001 vs. oeNC; ** P < 0.01 vs. oe-Myr-AKT1. C - K : In HGC-27 and MGC803 cells with AKT1 overexpression, LY294002 and shIRS-1 lentivirus were applied to assess the effects on cell proliferation (EdU) (C-E, ×100), invasion (Transwell) ( F , H , I , ×200), and migration (scratch) (G, J, K, ×40) ( n = 3 per group). Significance: ## P < 0.01, ### P < 0.001 vs. oeNC; * P < 0.05, ** P < 0.01, *** P < 0.001 vs. oe-Myr-AKT1

    Article Snippet: Fetal bovine serum (FBS, Beijing Cellmax Company, SA111.02); RPMI-1640 (Gibco, 42401042); DMEM (Dalian Meilun Biology, MA0212-2); LipofiterTM Transfection Reagent (Hanheng, HB-TRCF-1000); RIPA Lysis buffer (Beijing Solarbio Company, R0010); SDS Lysis Buffer (Beyotime, P0013G); Membrane Protein Extraction Kit (Solarbio, EX1500); Nuclear Protein Extraction Kit (Solarbio, EX1470); Notch1 Antibody (CST, 3447); Recombinant AKT1 Protein (Abcam, ab79792); Notch1-IC Antibody (CST, 4147); AKT1 Antibody (Abcam, ab238477); phospho-Akt substrate RXX(S*/T*) Antibody (CST, 9614); IRS-1 Antibody (Abcam, ab131487); Flag Antibody (Abcam, ab205606); E-cadherin Antibody(Abcam, ab231303); Snail1 Atibody(CST, 3879); Vimentin Antibody(Abcam, ab92547); Co-IP Kit (Thermo, 26149); Kinase Buffer (CST, 9802); ATP (CST, 9804); Recombinant human AKT1 protein (Active) (Abcam, ab62279); Recombinant Human NOTCH1 protein (hFc Tag) (Sinobiological, 10954-H02H); TRIzol (Ambion, USA, 15596026); PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) (TaKaRa, Japan, RR047A); TB Green ® Premix Ex Taq™ II (Tli RNaseH Plus) (TaKaRa, Japan, RR820A); PMSF (Beijing Solarbio Company, P0100); SDS-PAGE Gel Preparation Kit (Beijing Solarbio Company, P1200); ECL Ultra-Sensitive Luminol Reagent (Beijing Solarbio Company, PE0010); Pierce Co-IP Kit(Thermo Scientific, 26149), Protein A Agarose/Salmon Sperm DNA (Sigma-Aldrich, 16–157).

    Techniques: Western Blot, Expressing, Over Expression, Migration

    AKT1-Mediated Notch1 Phosphorylation Promotes the Growth of Gastric Cancer Cells In Vivo through the Regulation of IRS-1. A : Nude mice were injected with MGC803 cells overexpressing AKT1 alone or co-transfected with shIRS-1, with LY294002 treatment to monitor tumorigenesis. B : Statistical analysis of tumor volume at different time points ( n = 4 per group). C - J : IHC (×200) and Western blot analyzed EMT marker expression in tumor tissues ( n = 3 per group). K - M : Western blot assessed AKT1 expression and phosphorylation, with semi-quantitative analysis ( n = 3 per group). N - Q : Expression levels of IRS-1 and Notch1 were evaluated by IHC (×200) and Western blot ( n = 3 per group). I - S : Notch1-IC expression was detected using Western blot ( n = 3 per group). Significance: vs. oeNC group, ## P < 0.01, ### P < 0.001; vs. oe-Myr-AKT1 group, * P < 0.05, ** P < 0.01, *** P < 0.001

    Journal: Journal of Cancer Research and Clinical Oncology

    Article Title: AKT1-Mediated NOTCH1 phosphorylation promotes gastric cancer progression via targeted regulation of IRS-1 transcription

    doi: 10.1007/s00432-024-06039-z

    Figure Lengend Snippet: AKT1-Mediated Notch1 Phosphorylation Promotes the Growth of Gastric Cancer Cells In Vivo through the Regulation of IRS-1. A : Nude mice were injected with MGC803 cells overexpressing AKT1 alone or co-transfected with shIRS-1, with LY294002 treatment to monitor tumorigenesis. B : Statistical analysis of tumor volume at different time points ( n = 4 per group). C - J : IHC (×200) and Western blot analyzed EMT marker expression in tumor tissues ( n = 3 per group). K - M : Western blot assessed AKT1 expression and phosphorylation, with semi-quantitative analysis ( n = 3 per group). N - Q : Expression levels of IRS-1 and Notch1 were evaluated by IHC (×200) and Western blot ( n = 3 per group). I - S : Notch1-IC expression was detected using Western blot ( n = 3 per group). Significance: vs. oeNC group, ## P < 0.01, ### P < 0.001; vs. oe-Myr-AKT1 group, * P < 0.05, ** P < 0.01, *** P < 0.001

    Article Snippet: Fetal bovine serum (FBS, Beijing Cellmax Company, SA111.02); RPMI-1640 (Gibco, 42401042); DMEM (Dalian Meilun Biology, MA0212-2); LipofiterTM Transfection Reagent (Hanheng, HB-TRCF-1000); RIPA Lysis buffer (Beijing Solarbio Company, R0010); SDS Lysis Buffer (Beyotime, P0013G); Membrane Protein Extraction Kit (Solarbio, EX1500); Nuclear Protein Extraction Kit (Solarbio, EX1470); Notch1 Antibody (CST, 3447); Recombinant AKT1 Protein (Abcam, ab79792); Notch1-IC Antibody (CST, 4147); AKT1 Antibody (Abcam, ab238477); phospho-Akt substrate RXX(S*/T*) Antibody (CST, 9614); IRS-1 Antibody (Abcam, ab131487); Flag Antibody (Abcam, ab205606); E-cadherin Antibody(Abcam, ab231303); Snail1 Atibody(CST, 3879); Vimentin Antibody(Abcam, ab92547); Co-IP Kit (Thermo, 26149); Kinase Buffer (CST, 9802); ATP (CST, 9804); Recombinant human AKT1 protein (Active) (Abcam, ab62279); Recombinant Human NOTCH1 protein (hFc Tag) (Sinobiological, 10954-H02H); TRIzol (Ambion, USA, 15596026); PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) (TaKaRa, Japan, RR047A); TB Green ® Premix Ex Taq™ II (Tli RNaseH Plus) (TaKaRa, Japan, RR820A); PMSF (Beijing Solarbio Company, P0100); SDS-PAGE Gel Preparation Kit (Beijing Solarbio Company, P1200); ECL Ultra-Sensitive Luminol Reagent (Beijing Solarbio Company, PE0010); Pierce Co-IP Kit(Thermo Scientific, 26149), Protein A Agarose/Salmon Sperm DNA (Sigma-Aldrich, 16–157).

    Techniques: In Vivo, Injection, Transfection, Western Blot, Marker, Expressing

    A mechanistic illustration of AKT1 phosphorylating notch1-IC to regulate IRS-1 expression

    Journal: Journal of Cancer Research and Clinical Oncology

    Article Title: AKT1-Mediated NOTCH1 phosphorylation promotes gastric cancer progression via targeted regulation of IRS-1 transcription

    doi: 10.1007/s00432-024-06039-z

    Figure Lengend Snippet: A mechanistic illustration of AKT1 phosphorylating notch1-IC to regulate IRS-1 expression

    Article Snippet: Fetal bovine serum (FBS, Beijing Cellmax Company, SA111.02); RPMI-1640 (Gibco, 42401042); DMEM (Dalian Meilun Biology, MA0212-2); LipofiterTM Transfection Reagent (Hanheng, HB-TRCF-1000); RIPA Lysis buffer (Beijing Solarbio Company, R0010); SDS Lysis Buffer (Beyotime, P0013G); Membrane Protein Extraction Kit (Solarbio, EX1500); Nuclear Protein Extraction Kit (Solarbio, EX1470); Notch1 Antibody (CST, 3447); Recombinant AKT1 Protein (Abcam, ab79792); Notch1-IC Antibody (CST, 4147); AKT1 Antibody (Abcam, ab238477); phospho-Akt substrate RXX(S*/T*) Antibody (CST, 9614); IRS-1 Antibody (Abcam, ab131487); Flag Antibody (Abcam, ab205606); E-cadherin Antibody(Abcam, ab231303); Snail1 Atibody(CST, 3879); Vimentin Antibody(Abcam, ab92547); Co-IP Kit (Thermo, 26149); Kinase Buffer (CST, 9802); ATP (CST, 9804); Recombinant human AKT1 protein (Active) (Abcam, ab62279); Recombinant Human NOTCH1 protein (hFc Tag) (Sinobiological, 10954-H02H); TRIzol (Ambion, USA, 15596026); PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) (TaKaRa, Japan, RR047A); TB Green ® Premix Ex Taq™ II (Tli RNaseH Plus) (TaKaRa, Japan, RR820A); PMSF (Beijing Solarbio Company, P0100); SDS-PAGE Gel Preparation Kit (Beijing Solarbio Company, P1200); ECL Ultra-Sensitive Luminol Reagent (Beijing Solarbio Company, PE0010); Pierce Co-IP Kit(Thermo Scientific, 26149), Protein A Agarose/Salmon Sperm DNA (Sigma-Aldrich, 16–157).

    Techniques: Expressing

    ( a–c ) Representative triple stain images for N1ICD, Notch1, and Jag2 mRNA in sebaceous glands (SGs) from mice (n=5 each) treated with aRW, 3 d post-treatment. ( a ) Four channels showing N1ICD, N1 ISH, J2 ISH, and DAPI. ( b ) Two channels showing N1 ISH and J2 ISH. ( c ) One channel showing N1ICD. ( d-f ) Representative triple stain images for N1ICD, Notch1, and Jag2 mRNA in SGs from mice (n=5 each) treated with aJ1, 3 d post-treatment. ( d ) Four channels showing N1ICD, N1 ISH, J2 ISH, and DAPI. ( e ) Two channels showing N1 ISH and J2 ISH. ( f ) One channel showing N1ICD. ( g ) Quantification of the percentage of N1ICD positive (N1ICD+) basal stem cells in SGs from mice (n=5 each) treated with aRW, aJ1, and aJ2, 3 d post-treatment. Percentage was calculated by dividing the number of N1ICD+ basal stem cells by the total number of basal stem cells in each SG. ( h–j ) Representative triple stain images for N1ICD, Notch1, and Jag2 mRNA in SGs from mice (n=5 each) treated with aJ2, 3 d post-treatment. ( h ) Four channels showing N1ICD, N1 ISH, J2 ISH, and DAPI. ( i ) Two channels showing N1 ISH and J2 ISH. ( j ) One channel showing N1ICD. ( k ) Quantification of what percentage of the N1ICD+ basal stem cells express both N1 ISH and J2 ISH, only N1 ISH, or only J2 ISH. ( g and k ) Total n of SGs quantified per treatment: aRW=13, aJ1=15, aJ2=15. Error bars represent SEM. Scale bars are 25 μm. Figure 2—source data 1. Source data for .

    Journal: eLife

    Article Title: The Jag2/Notch1 signaling axis promotes sebaceous gland differentiation and controls progenitor proliferation

    doi: 10.7554/eLife.98747

    Figure Lengend Snippet: ( a–c ) Representative triple stain images for N1ICD, Notch1, and Jag2 mRNA in sebaceous glands (SGs) from mice (n=5 each) treated with aRW, 3 d post-treatment. ( a ) Four channels showing N1ICD, N1 ISH, J2 ISH, and DAPI. ( b ) Two channels showing N1 ISH and J2 ISH. ( c ) One channel showing N1ICD. ( d-f ) Representative triple stain images for N1ICD, Notch1, and Jag2 mRNA in SGs from mice (n=5 each) treated with aJ1, 3 d post-treatment. ( d ) Four channels showing N1ICD, N1 ISH, J2 ISH, and DAPI. ( e ) Two channels showing N1 ISH and J2 ISH. ( f ) One channel showing N1ICD. ( g ) Quantification of the percentage of N1ICD positive (N1ICD+) basal stem cells in SGs from mice (n=5 each) treated with aRW, aJ1, and aJ2, 3 d post-treatment. Percentage was calculated by dividing the number of N1ICD+ basal stem cells by the total number of basal stem cells in each SG. ( h–j ) Representative triple stain images for N1ICD, Notch1, and Jag2 mRNA in SGs from mice (n=5 each) treated with aJ2, 3 d post-treatment. ( h ) Four channels showing N1ICD, N1 ISH, J2 ISH, and DAPI. ( i ) Two channels showing N1 ISH and J2 ISH. ( j ) One channel showing N1ICD. ( k ) Quantification of what percentage of the N1ICD+ basal stem cells express both N1 ISH and J2 ISH, only N1 ISH, or only J2 ISH. ( g and k ) Total n of SGs quantified per treatment: aRW=13, aJ1=15, aJ2=15. Error bars represent SEM. Scale bars are 25 μm. Figure 2—source data 1. Source data for .

    Article Snippet: Antibody , Anti-Notch1 (Human monoclonal) , Genentech , , Inhibiting antibody (5 mg/kg).

    Techniques: Staining

    (Related to ). ( a–c ) Quantification of the location of N1ICD+ basal stem cells in sebaceous glands (SGs) from mice (n=5 each) treated with aRW ( a ), aJ1 ( b ), and aJ2 ( c ), 3 d post-treatment. The SG was divided into five sections along the proximal-distal axis. The number of N1ICD+ basal stem cells in each section was divided by the total number of N1ICD+ basal stem cells in each SG. Total n of SGs quantified per treatment: aRW=13, aJ1=15, aJ2=15. ( d ) Quantification of what percentage of the total basal stem cells (N1ICD+ and N1ICD-) express both N1 ISH and J2 ISH. Total n of SGs quantified per treatment: aRW=13, aJ1=15, aJ2=15. ( e,f ) Representative triple stain images for N1ICD, Notch1, and Jag2 mRNA in SGs from mice (n=5 each) treated with ( e ) aRW and ( f ) aJ2, 3 d post-treatment. ( g ) Quantification of the width of the interfollicular epidermis (IFE), 7 d after treatment with aRW, aJ1, aJ2, and aJ1J2. p-values: aJ1=0.528, aJ2=0.885, aJ1J2=0.840. Total n of SGs quantified per treatment: aRW=26, aJ1=27, aJ2=27, aJ1J2=22. ( h ) Quantification of the width of the adipocyte layer, 7 d after treatment with aRW, aJ1, aJ2, and aJ1J2. p-values: aJ1=0.890, aJ2=0.948, aJ1J2=0.374. Total n of SGs quantified per treatment: aRW=13, aJ1=13, aJ2=15, aJ1J2=15. ( i,j ) Representative hematoxylin and eosin (H&E) images of SGs from mice (n=5 each) treated with aRW ( i ) and aJ2 ( j ), 7 d post-treatment. Student’s t-test used for statistical analysis. Error bars represent SEM. Scale bars are 100 μm. Figure 2—figure supplement 1—source data 1. Source data for .

    Journal: eLife

    Article Title: The Jag2/Notch1 signaling axis promotes sebaceous gland differentiation and controls progenitor proliferation

    doi: 10.7554/eLife.98747

    Figure Lengend Snippet: (Related to ). ( a–c ) Quantification of the location of N1ICD+ basal stem cells in sebaceous glands (SGs) from mice (n=5 each) treated with aRW ( a ), aJ1 ( b ), and aJ2 ( c ), 3 d post-treatment. The SG was divided into five sections along the proximal-distal axis. The number of N1ICD+ basal stem cells in each section was divided by the total number of N1ICD+ basal stem cells in each SG. Total n of SGs quantified per treatment: aRW=13, aJ1=15, aJ2=15. ( d ) Quantification of what percentage of the total basal stem cells (N1ICD+ and N1ICD-) express both N1 ISH and J2 ISH. Total n of SGs quantified per treatment: aRW=13, aJ1=15, aJ2=15. ( e,f ) Representative triple stain images for N1ICD, Notch1, and Jag2 mRNA in SGs from mice (n=5 each) treated with ( e ) aRW and ( f ) aJ2, 3 d post-treatment. ( g ) Quantification of the width of the interfollicular epidermis (IFE), 7 d after treatment with aRW, aJ1, aJ2, and aJ1J2. p-values: aJ1=0.528, aJ2=0.885, aJ1J2=0.840. Total n of SGs quantified per treatment: aRW=26, aJ1=27, aJ2=27, aJ1J2=22. ( h ) Quantification of the width of the adipocyte layer, 7 d after treatment with aRW, aJ1, aJ2, and aJ1J2. p-values: aJ1=0.890, aJ2=0.948, aJ1J2=0.374. Total n of SGs quantified per treatment: aRW=13, aJ1=13, aJ2=15, aJ1J2=15. ( i,j ) Representative hematoxylin and eosin (H&E) images of SGs from mice (n=5 each) treated with aRW ( i ) and aJ2 ( j ), 7 d post-treatment. Student’s t-test used for statistical analysis. Error bars represent SEM. Scale bars are 100 μm. Figure 2—figure supplement 1—source data 1. Source data for .

    Article Snippet: Antibody , Anti-Notch1 (Human monoclonal) , Genentech , , Inhibiting antibody (5 mg/kg).

    Techniques: Staining

    Journal: eLife

    Article Title: The Jag2/Notch1 signaling axis promotes sebaceous gland differentiation and controls progenitor proliferation

    doi: 10.7554/eLife.98747

    Figure Lengend Snippet:

    Article Snippet: Antibody , Anti-Notch1 (Human monoclonal) , Genentech , , Inhibiting antibody (5 mg/kg).

    Techniques: Control, Staining, Sequencing, RNAscope, Multiplex Assay