Journal: medRxiv

Article Title: CRABP2 Upregulation in Perichondral Stem Cells is Associated with Microtia

doi: 10.64898/2025.12.18.25342452

Figure Lengend Snippet: (A) Boxplot shows the RA responsiveness score measured by three different gene sets. (B) Dot plot shows the top 10 active regulons predicted by pyscenic in the COL1 + HES1 + perichondral stem cells from normal and microtia samples respectively. (C) Gene ontology enrichment results of genes up-regulated in COL1 + HES1 + perichondral stem cells from normal samples (left) and in microtia samples (right). (D) Expression level of RA associated genes in COL1 + HES1 + perichondral stem cells from normal samples (left) and in microtia samples (right). ***P<0.001, Student’s t test.

Article Snippet: The following primary antibodies and dilutions were used: COL-1 (1:150, 67288-1-Ig, Proteintech, China), CoraLite 594-conjugated CRABP2 (1:75, CL594-10225, Proteintech, China), and HES1 (1:200, 11988, CST, United States).

Techniques: Expressing

Journal: medRxiv

Article Title: CRABP2 Upregulation in Perichondral Stem Cells is Associated with Microtia

doi: 10.64898/2025.12.18.25342452

Figure Lengend Snippet: (A) Representative immunofluorescence staining images of auricular cartilage slices from microtia and normal samples. Scale bar for large images: 150 µm. Scale bar for insets: 50 µm. (B) Line intensity profiles of DAPI, anti-COL1, anti-CRABP2 and anti-HES1 immunostainings. (C) Boxplot shows the quantification result of CRABP2 + COL1 + HES1 + proportions among COL1 + HES1 + perichondral stem cells in normal and microtia samples. *P<0.05, Student’s t test.

Article Snippet: The following primary antibodies and dilutions were used: COL-1 (1:150, 67288-1-Ig, Proteintech, China), CoraLite 594-conjugated CRABP2 (1:75, CL594-10225, Proteintech, China), and HES1 (1:200, 11988, CST, United States).

Techniques: Immunofluorescence, Staining

Journal: Aging and Disease

Article Title: Mash1-dependent Notch Signaling Pathway Regulates GABAergic Neuron-Like Differentiation from Bone Marrow-Derived Mesenchymal Stem Cells

doi: 10.14336/AD.2016.1018

Figure Lengend Snippet: Primer sequences for Notch signaling and GABAergic neuron marker

Article Snippet: For RBPJ or Hes1 silencing, BMSCs which reached approximately 50% confluence in a 12-well plate were replaced with Polybrene (Santa Cruz Biotechnology, Inc., sc-134220, CA, USA) media mixture, followed by infection with RBPJ (Santa Cruz Biotechnology, Inc., sc-270318-v, CA, USA) or Hes1 (Santa Cruz Biotechnology, Inc., sc-270146-v, CA, USA) shRNA lentiviral particles (these primers were provided by the company) overnight (37°C, 5% CO 2 ).

Techniques: Sequencing

Journal: Aging and Disease

Article Title: Mash1-dependent Notch Signaling Pathway Regulates GABAergic Neuron-Like Differentiation from Bone Marrow-Derived Mesenchymal Stem Cells

doi: 10.14336/AD.2016.1018

Figure Lengend Snippet: A ) Protein bands of Notch signaling (RBPJ, Hes1 and Mash1) visualized through western blotting of BMSCs that were treated by DAPT (DAPT+BMSCs), or engineered by Hes1 shRNA (Hes1-BMSCs), pGC-FU-Mash1 plasmid (Mash1+BMSCs), RBPJ shRNA (RBPJ-BMSCs) and copGFP Control (Lv-con-BMSCs). B ) The protein level based on the ratio of Gauss Model Trace exhibited the Notch signaling (RBPJ, Hes1 and Mash1) changes in the DAPT+ BMSCs. C ) The protein level based on the ratio of Gauss Model Trace exhibited genetically engineered BMSCs. Error bars in bar graphs display standard deviation (SD). * P = 0.032, < 0.05, ** P = 0.0006, < 0.01.

Article Snippet: For RBPJ or Hes1 silencing, BMSCs which reached approximately 50% confluence in a 12-well plate were replaced with Polybrene (Santa Cruz Biotechnology, Inc., sc-134220, CA, USA) media mixture, followed by infection with RBPJ (Santa Cruz Biotechnology, Inc., sc-270318-v, CA, USA) or Hes1 (Santa Cruz Biotechnology, Inc., sc-270146-v, CA, USA) shRNA lentiviral particles (these primers were provided by the company) overnight (37°C, 5% CO 2 ).

Techniques: Western Blot, shRNA, Plasmid Preparation, Control, Standard Deviation

Journal: World Journal of Gastroenterology

Article Title: Downregulation of Hes1 expression in experimental biliary atresia and its effects on bile duct structure

doi: 10.3748/wjg.v24.i29.3260

Figure Lengend Snippet: Primers used in the experiments

Article Snippet: To study the interruption of gene expression, such as interruption of the Notch signaling pathway target molecule Hes1, in the IBEC 3D cell culture, Hes1 siRNA RNA and control siRNA purchased from Santa Cruz Biotechnology, Inc., were used in the cell culture and transfected into cells using Lipofectamine ® RNAiMAX Reagent (Life Technologies, 13778) following the manufacturer’s instructions.

Techniques:

Journal: World Journal of Gastroenterology

Article Title: Downregulation of Hes1 expression in experimental biliary atresia and its effects on bile duct structure

doi: 10.3748/wjg.v24.i29.3260

Figure Lengend Snippet: Expression levels of Notch signaling components in the liver of rhesus rotavirus-inoculated mice. The mRNA levels of Notch signaling components, including Jag1, Dll1, Notch1, Notch2 and Hes1, were detected using quantitative real-time PCR with appropriate primers at day 7 and day 14 after RRV inoculation. A: The relative expression levels compared with the normal control are shown for day 7 ( a P < 0.05; b P < 0.01; n = 6 in the control group and n = 10 in the RRV group); B: day 14 ( c P < 0.001); C: The levels of the Hes1 protein and the expression of the transcriptional coregulator RBP-Jκ were detected by Western blot ( n = 2 or 3 in each group; the proteins were mixed and loaded).

Article Snippet: To study the interruption of gene expression, such as interruption of the Notch signaling pathway target molecule Hes1, in the IBEC 3D cell culture, Hes1 siRNA RNA and control siRNA purchased from Santa Cruz Biotechnology, Inc., were used in the cell culture and transfected into cells using Lipofectamine ® RNAiMAX Reagent (Life Technologies, 13778) following the manufacturer’s instructions.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Western Blot

Journal: World Journal of Gastroenterology

Article Title: Downregulation of Hes1 expression in experimental biliary atresia and its effects on bile duct structure

doi: 10.3748/wjg.v24.i29.3260

Figure Lengend Snippet: Hes1 siRNA in 3D biliary epithelial cell culture. Biliary epithelial cells (BECs) were cultured in either normal growth medium (N. medium) or in matrix + collagen mixed gel (detailed components are given in the Materials and Methods section). The cells were cultured without a transfection reagent (Cont.), with Lipofectamine (Lipo) only, with irrelevant control siRNA (Cont. siRNA) or with Hes1 siRNA for seven days, then fixed and stained with anti-CK19, γ-GT and AFP antibodies, and finally counter-stained with Alexa Fluor ® 488. Photographs of the cells were taken with a Leica DMi8 inverted fluorescence microscope, and the groups were then compared. The experiment was duplicated, and one set of representative results is shown here.

Article Snippet: To study the interruption of gene expression, such as interruption of the Notch signaling pathway target molecule Hes1, in the IBEC 3D cell culture, Hes1 siRNA RNA and control siRNA purchased from Santa Cruz Biotechnology, Inc., were used in the cell culture and transfected into cells using Lipofectamine ® RNAiMAX Reagent (Life Technologies, 13778) following the manufacturer’s instructions.

Techniques: Cell Culture, Transfection, Control, Staining, Fluorescence, Microscopy

Journal: Naunyn-Schmiedeberg's Archives of Pharmacology

Article Title: Tanshinol ameliorates imiquimod-induced psoriasis by inhibiting M1 macrophage polarization through suppression of the notch signaling pathway

doi: 10.1007/s00210-024-03166-9

Figure Lengend Snippet: Effect of TAN on Notch signaling pathway in skin lesion tissues of psoriasis mice. (A) The protein expression of Notch1, Hey1, and Hes1 in dorsal skin tissues. (B) Immunofluorescence co-staining of F4/80 + Notch1 (20×, Scale bar, 50 μm). Data were represented as mean ± SEM. Compared with control group, ** P < 0.01; compared with model group, ## P < 0.01

Article Snippet: Total proteins were electrophoresed on SDS-PAGE polyacrylamide gels and transferred to PVDF membranes, closed with 5% skimmed milk, and then incubated overnight at 4°C with primary antibodies [Notch1 (cat no: A16673), Hey1 (cat no: A16110), Hes1 (cat no: A0925), Abclonal, Wuhan], secondary antibodies Goat Anti-Rabbit IgG (H + L) HRP (cat no: S0001, Affbiotech, Suzhou) incubated at room temperature for 1 h, developed by ECL, and the bands were exposed with Fluorescence Image Analysis System Software V2.0 (Tanon, Shanghai), and the results were scanned by Gel-Pro analyzer4 software and expressed as the integrated optical density (IOD) of the target protein.

Techniques: Expressing, Immunofluorescence, Staining, Control

Journal: Naunyn-Schmiedeberg's Archives of Pharmacology

Article Title: Tanshinol ameliorates imiquimod-induced psoriasis by inhibiting M1 macrophage polarization through suppression of the notch signaling pathway

doi: 10.1007/s00210-024-03166-9

Figure Lengend Snippet: Effect of TAN on skin lesion phenotype and macrophage polarization in psoriasis mice through inhibition of the Notch signaling pathway. (A) Photograph of the skin on the back of a mouse. (B) PASI score, epidermal thickness, and ear thickness. (C) Histopathologic picture of the skin of the back (HE, Scale bar, 200 μm and 50 μm). (D) The expression of ki67, K6, F4/80 + iNOS, and F4/80 + CD206 in dorsal skin tissues. (E) Immunofluorescence staining of ki67 and K6 (20×, Scale bar, 50 μm). (F) Immunofluorescence co-staining of F4/80 + iNOS and F4/80 + CD206 (20×, Scale bar, 50 μm). (G-H) The protein expression of Notch1, Hey1, and Hes1 in dorsal skin tissues. (I-J) The expression of MCP-1 and MIF in dorsal skin tissues (immunohistochemical staining, 20×, Scale bar, 50 μm). (K) The mRNA expression of iNOS, TNFα, IL-6, IL-20, MIF, MCP-1, IL-4 in dorsal skin tissues. Data were represented as mean ± SEM. Compared with control group, * P < 0.05, ** P < 0.01; compared with model group, # P < 0.05, ## P < 0.01; compared with the TAN-H group, Δ P < 0.05, ΔΔ P < 0.01

Article Snippet: Total proteins were electrophoresed on SDS-PAGE polyacrylamide gels and transferred to PVDF membranes, closed with 5% skimmed milk, and then incubated overnight at 4°C with primary antibodies [Notch1 (cat no: A16673), Hey1 (cat no: A16110), Hes1 (cat no: A0925), Abclonal, Wuhan], secondary antibodies Goat Anti-Rabbit IgG (H + L) HRP (cat no: S0001, Affbiotech, Suzhou) incubated at room temperature for 1 h, developed by ECL, and the bands were exposed with Fluorescence Image Analysis System Software V2.0 (Tanon, Shanghai), and the results were scanned by Gel-Pro analyzer4 software and expressed as the integrated optical density (IOD) of the target protein.

Techniques: Inhibition, Expressing, Immunofluorescence, Staining, Immunohistochemical staining, Control

Journal: Naunyn-Schmiedeberg's Archives of Pharmacology

Article Title: Tanshinol ameliorates imiquimod-induced psoriasis by inhibiting M1 macrophage polarization through suppression of the notch signaling pathway

doi: 10.1007/s00210-024-03166-9

Figure Lengend Snippet: Effect of TAN on macrophage polarization, cytokine secretion and Notch pathway. (A) Flow cytogram of F4/80 + CD86 and F4/80 + CD163. (B) Cell viability. (C) Levels of F4/80 + CD86 and F4/80 + CD163. (D) Levels of iNOS, IL-6, IL-20, MIF, MCP-1, and IL-4. (E) The protein expression of Notch1, Hey1, and Hes1. Data were represented as mean ± SEM. Compared with control group, ** P < 0.01; compared with M1 group, # P < 0.05, ## P < 0.01

Article Snippet: Total proteins were electrophoresed on SDS-PAGE polyacrylamide gels and transferred to PVDF membranes, closed with 5% skimmed milk, and then incubated overnight at 4°C with primary antibodies [Notch1 (cat no: A16673), Hey1 (cat no: A16110), Hes1 (cat no: A0925), Abclonal, Wuhan], secondary antibodies Goat Anti-Rabbit IgG (H + L) HRP (cat no: S0001, Affbiotech, Suzhou) incubated at room temperature for 1 h, developed by ECL, and the bands were exposed with Fluorescence Image Analysis System Software V2.0 (Tanon, Shanghai), and the results were scanned by Gel-Pro analyzer4 software and expressed as the integrated optical density (IOD) of the target protein.

Techniques: Expressing, Control

Journal: Naunyn-Schmiedeberg's Archives of Pharmacology

Article Title: Tanshinol ameliorates imiquimod-induced psoriasis by inhibiting M1 macrophage polarization through suppression of the notch signaling pathway

doi: 10.1007/s00210-024-03166-9

Figure Lengend Snippet: TAN regulates macrophage polarization and cytokine secretion through the Notch pathway. (A) Flow cytogram of F4/80 + CD86 and F4/80 + CD163. (B) Levels of F4/80 + CD86 and F4/80 + CD163. (C) . Levels of iNOS, IL-6, IL-20, MIF, MCP-1, and IL-4. (D) The protein expression of Notch1, Hey1, and Hes1. Data were represented as mean ± SEM. Compared with M1 group, * P < 0.05, ** P < 0.01; compared with M1 + TAN-H group, # P < 0.05, ## P < 0.01; compared with M1 + Jagged1 group, Δ P < 0.05, ΔΔ P < 0.01

Article Snippet: Total proteins were electrophoresed on SDS-PAGE polyacrylamide gels and transferred to PVDF membranes, closed with 5% skimmed milk, and then incubated overnight at 4°C with primary antibodies [Notch1 (cat no: A16673), Hey1 (cat no: A16110), Hes1 (cat no: A0925), Abclonal, Wuhan], secondary antibodies Goat Anti-Rabbit IgG (H + L) HRP (cat no: S0001, Affbiotech, Suzhou) incubated at room temperature for 1 h, developed by ECL, and the bands were exposed with Fluorescence Image Analysis System Software V2.0 (Tanon, Shanghai), and the results were scanned by Gel-Pro analyzer4 software and expressed as the integrated optical density (IOD) of the target protein.

Techniques: Expressing