notch ic  (R&D Systems)

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: The overexpression human Flag-Myr-AKT1 and Notch-IC lentiviral vectors were purchased from Shanghai Hanheng Bio-Tech Co., Ltd.

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

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: The overexpression human Flag-Myr-AKT1 and Notch-IC lentiviral vectors were purchased from Shanghai Hanheng Bio-Tech Co., Ltd.

Techniques: Western Blot, Expressing, Over Expression, Migration

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: The overexpression human Flag-Myr-AKT1 and Notch-IC lentiviral vectors were purchased from Shanghai Hanheng Bio-Tech Co., Ltd.

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

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: The overexpression human Flag-Myr-AKT1 and Notch-IC lentiviral vectors were purchased from Shanghai Hanheng Bio-Tech Co., Ltd.

Techniques: Expressing

Journal: Developmental biology

Article Title: Origin of the brush cell lineage in the mouse intestinal epithelium

doi: 10.1016/j.ydbio.2011.12.009

Figure Lengend Snippet: Recombination efficiency before and after Tamoxifen treatment.

Article Snippet: For the Notch-IC clone study, a female homozygous Rosa26 floxed-STOP-Notch-IC-IRES-EGFP mouse ( Murtaugh et al., 2003 ; The Jackson Laboratory, 008159), was mated with a male homozygous Rosa26 CreERT2 mouse to generate F 1 Rosa26CreERT2/fl oxed-STOP-Notch-IC-IRES-EGFP mice.

Techniques:

Journal: Developmental biology

Article Title: Origin of the brush cell lineage in the mouse intestinal epithelium

doi: 10.1016/j.ydbio.2011.12.009

Figure Lengend Snippet: Lineage tracing with Atoh1-Cre reporter mice indicates that brush lineage precursors transiently express Atoh1. (A–D) Optical sections of villi from Atoh1-Cre;Rosa26-LacZ lineage tracing mice showing, as expected, that Atoh1 expression has occurred in mucous (arrows) and enteroendocrine (arrow heads) lineage cells or their precursors. Evidence for the Paneth cell lineage is shown in Fig. 16D. In contrast, the vast majority of brush lineage cells (boxed) were negative, but a small number were β-gal+ (C) indicating Atoh1 expression occurred at some point in their past. (D) Low magnification image (X-gal stained) of a rare large stem cell derived clone on the villus. (E–H) Similar results were obtained from Atoh1-CreERT2; Rosa26-LacZ mice, except that β-gal+ brush cells (G) were more frequent, and importantly, that clones containing multiple columnar cells (H) were consistently seen indicating that a low level of transient Atoh1 expression occurs in early columnar lineage precursors, most likely the daughter of Mix (DOM) progenitors.

Article Snippet: For the Notch-IC clone study, a female homozygous Rosa26 floxed-STOP-Notch-IC-IRES-EGFP mouse ( Murtaugh et al., 2003 ; The Jackson Laboratory, 008159), was mated with a male homozygous Rosa26 CreERT2 mouse to generate F 1 Rosa26CreERT2/fl oxed-STOP-Notch-IC-IRES-EGFP mice.

Techniques: Expressing, Staining, Derivative Assay, Clone Assay

Journal: Developmental biology

Article Title: Origin of the brush cell lineage in the mouse intestinal epithelium

doi: 10.1016/j.ydbio.2011.12.009

Figure Lengend Snippet: The nuclei of some immature Gfi1b expressing cells in the lower crypt stain weakly with Hes1 antibody. (A, B) The nuclei of most brush lineage cells (boxed) do not stain with Hes1 antibody. (A) A CD-1 mouse crypt with two Gfi1b+ Hes1− brush cell lineage nuclei (the Gfi1b staining of Paneth granules in the crypt base is nonspecific). (B) A tangential optical section of a Gfi1bEGFP/+ crypt containing 5 EGFPGfi1b+ cells illustrating the increasing gradient of differentiation as brush lineage cells migrate up the crypt from their origin in the COD. The cells display an increasing gradient of cell size and EGFPGfi1b content. Only the small, immature cell in the crypt base is weakly positive for Hes1 (barbed arrow). (C) A weakly Gfi1b+ Hes1+ brush lineage cell in the base and a Gfi1b+ Hes1− brush lineage cell in the top of a crypt, isolated from an Atoh1 fl/fl; Rosa26 Cre/+ mouse after Tamoxifen treatment. (D) Optical section of a crypt from an Atoh1-Cre;Rosa26-LacZ mouse showing Atoh1 expression in Paneth cells (barbed arrows). Paneth cell granules are visualized by UEA-I staining.

Article Snippet: For the Notch-IC clone study, a female homozygous Rosa26 floxed-STOP-Notch-IC-IRES-EGFP mouse ( Murtaugh et al., 2003 ; The Jackson Laboratory, 008159), was mated with a male homozygous Rosa26 CreERT2 mouse to generate F 1 Rosa26CreERT2/fl oxed-STOP-Notch-IC-IRES-EGFP mice.

Techniques: Expressing, Staining, Isolation

Journal: Developmental biology

Article Title: Origin of the brush cell lineage in the mouse intestinal epithelium

doi: 10.1016/j.ydbio.2011.12.009

Figure Lengend Snippet: Fraction of each cell type expressing the reporter following Atoh1-Cre lineage tracing.

Article Snippet: For the Notch-IC clone study, a female homozygous Rosa26 floxed-STOP-Notch-IC-IRES-EGFP mouse ( Murtaugh et al., 2003 ; The Jackson Laboratory, 008159), was mated with a male homozygous Rosa26 CreERT2 mouse to generate F 1 Rosa26CreERT2/fl oxed-STOP-Notch-IC-IRES-EGFP mice.

Techniques: Expressing

Journal: Developmental biology

Article Title: Origin of the brush cell lineage in the mouse intestinal epithelium

doi: 10.1016/j.ydbio.2011.12.009

Figure Lengend Snippet: Cell type counts after conditional Atoh1 deletion in Atoh1 fl/+ control and Atoh1 fl/fl experimental mice.

Article Snippet: For the Notch-IC clone study, a female homozygous Rosa26 floxed-STOP-Notch-IC-IRES-EGFP mouse ( Murtaugh et al., 2003 ; The Jackson Laboratory, 008159), was mated with a male homozygous Rosa26 CreERT2 mouse to generate F 1 Rosa26CreERT2/fl oxed-STOP-Notch-IC-IRES-EGFP mice.

Techniques: Control

Journal: Developmental biology

Article Title: Origin of the brush cell lineage in the mouse intestinal epithelium

doi: 10.1016/j.ydbio.2011.12.009

Figure Lengend Snippet: Counts of the number of Gfi1b expressing cells per crypt after conditional Atoh1 deletion in Atoh1 fl/+ control and Atoh1 fl/fl experimental mice.

Article Snippet: For the Notch-IC clone study, a female homozygous Rosa26 floxed-STOP-Notch-IC-IRES-EGFP mouse ( Murtaugh et al., 2003 ; The Jackson Laboratory, 008159), was mated with a male homozygous Rosa26 CreERT2 mouse to generate F 1 Rosa26CreERT2/fl oxed-STOP-Notch-IC-IRES-EGFP mice.

Techniques: Expressing, Control