Journal: iScience

Article Title: Spatiotemporally organized immunomodulatory response to SARS-CoV-2 virus in primary human broncho-alveolar epithelia

doi: 10.1016/j.isci.2023.107374

Figure Lengend Snippet:

Article Snippet: Anti-CSF3 AF594 (Clone CSF3/900) , Novus , CatNBP2-47934AF594 RRID: AB_2933966.

Techniques: Bacteria, Virus, Variant Assay, Recombinant, Red Blood Cell Lysis, Software, Membrane, Cell Culture

Journal: International journal of molecular sciences

Article Title: A NOTCH1 Mutation Found in a Newly Established Ovarian Cancer Cell Line (FDOVL) Promotes Lymph Node Metastasis in Ovarian Cancer.

doi: 10.3390/ijms24065091

Figure Lengend Snippet: Figure 3. CSF3 is determined as the downstream effector of NOTCH1-p.C720fs mutation on epithelial- mesenchymal transition. (A,B) CXCL1, CXCL2, CXCL3, CXCL8, CSF3 and IL1B were determined

Article Snippet: Similarly, immunohistochemistry (IHC) was conducted to assess the expression of Ki67 (1:200, AF0198, Affinity, San Francisco, CA, USA), CyclinD1 (1:250, ab134175, Abcam), Hes1 (1:200, ab108937, Abcam), N-cadherin (1:200, #13116, Cell Signaling Technology, Danvers, MA, USA), vimentin (1:200, #5741, Cell Signaling Technology) and CSF3 (1:200, 17185-1-AP, Proteintech) in the tumors extracted from mice.

Techniques: Mutagenesis

Journal: International journal of molecular sciences

Article Title: A NOTCH1 Mutation Found in a Newly Established Ovarian Cancer Cell Line (FDOVL) Promotes Lymph Node Metastasis in Ovarian Cancer.

doi: 10.3390/ijms24065091

Figure Lengend Snippet: Figure 5. NOTCH1 mutations are more prominent in metastatic lymph nodes of ovarian cancer than in other implant metastatic sites. (A,B) Ten paired abdominal implant lesions and metastatic lymph nodes of ovarian cancer were subjected to WES and IHC detection to assess the difference of driver genes and the expression of downstream CSF3 in implant lesions and metastatic lymph nodes. (A) The heat map of driver genes in 10 paired abdominal implant lesions and metastatic lymph nodes of ovarian cancer. (B) The expression level of CSF3 in lymph nodes harboring mutated NOTCH1 was significantly higher than that with wild-type NOTCH1. The representative image of scoring criteria on the expression of CSF3: “no staining”, “weak staining”, and “strong staining”, with the quantitative analysis of H score shown on the right. * p < 0.05.

Article Snippet: Similarly, immunohistochemistry (IHC) was conducted to assess the expression of Ki67 (1:200, AF0198, Affinity, San Francisco, CA, USA), CyclinD1 (1:250, ab134175, Abcam), Hes1 (1:200, ab108937, Abcam), N-cadherin (1:200, #13116, Cell Signaling Technology, Danvers, MA, USA), vimentin (1:200, #5741, Cell Signaling Technology) and CSF3 (1:200, 17185-1-AP, Proteintech) in the tumors extracted from mice.

Techniques: Expressing, Staining

Journal: American Journal of Cancer Research

Article Title: CHPF promotes malignancy of breast cancer cells by modifying syndecan-4 and the tumor microenvironment

doi:

Figure Lengend Snippet: CHPF promotes tumor growth, metastasis, and MDSC accumulation in tumor tissue. Overexpression of Chpf increases tumor weight (A) and lung metastasis (B) of 4T1 tumor model (n = 6; *P < 0.05). Images of dissected tumors and lungs are shown on the left. Arrows indicate lung tumor nodules. Scale bars: 1.0 cm. (C) Cytokine and chemokine levels in 4T1 tumor tissues. Tumor tissue lysate (100 mg) is analysed using multiplex immunoassay. *P < 0.05, **P < 0.01. (D) Chpf increases infiltrated MDSC numbers in tumor tissue. Representative images of gating Gr1 high (Gr1h) and Gr1 intermediate (Gr1int) MDSCs are shown on the right. The mean ± SD are shown on the left (n = 7). *P < 0.05, **P < 0.01. (E) Immunostaining of F4/80 and G-CSF in 4T1 tumor sections. (F) Immunostaining of Ly6G and G-CSF in 4T1 tumor sections. Amplified images are shown at the bottom right of each image. Scale bars: 20 μm.

Article Snippet: G-CSF antibody (bs-1023R) was purchased from Bioss Inc.

Techniques: Over Expression, Multiplex Assay, Immunostaining, Amplification

Journal: American Journal of Cancer Research

Article Title: CHPF promotes malignancy of breast cancer cells by modifying syndecan-4 and the tumor microenvironment

doi:

Figure Lengend Snippet: CHPF regulates G-CSF protein levels in breast cancer cells. A. mRNA levels of CHPF and G-CSF in Chpf-overexpressed and CHPF-silenced breast cancer cells. ***P < 0.001 compared to control groups. B. Protein levels of G-CSF in Chpf-overexpressed and CHPF-silenced breast cancer cells. C. Co-localisation of cell surface CS and G-CSF on 4T1 cells. Mock, CHPF clone, and β-D-xylopyranoside (β-XP)-treated cells were stained with CS56 antibody (green) and G-CSF antibody (red). Amplified images are shown on the right. Dashed circles indicate the area of the nucleus. Arrows point the co-localised (yellow) spots on the cells. Scale bars: 20 μm. D. Co-culture of MDSCs and 4T1 cells induced 4T1 cell invasion. Mock or Chpf-overexpressed cells in the transwell were co-cultured with or without freshly isolated MDSCs and incubated in 5% FBS-DMEM for 24 h. Invasive cells were counted and the representative images are shown on the right. Data are expressed as the mean ± SD from three independent experiments. ***P < 0.001.

Article Snippet: G-CSF antibody (bs-1023R) was purchased from Bioss Inc.

Techniques: Staining, Amplification, Co-Culture Assay, Cell Culture, Isolation, Incubation

Journal: American Journal of Cancer Research

Article Title: CHPF promotes malignancy of breast cancer cells by modifying syndecan-4 and the tumor microenvironment

doi:

Figure Lengend Snippet: Syndecan-4 is involved in CHPF-mediated breast cancer malignancy. A. Immunoprecipitation (IP) of 4T1 cell lysate using CS56 antibody. The immunoprecipitated protein is separated using 8% SDS-PAGE and visualised using stain-free technology (Bio-Rad). The red arrowheads indicate the CHPF-enhanced proteins after IP. The black arrowhead indicates the heavy chains of the antibody. B. CHPF modifies Syndecan-4 (SDC4). Western blots of SDC4 in mock or Chpf-overexpressed cell lysates. Chondroitinase ABC (ChABC) was used to digest CS in the cell lysate. C. Relative mRNA expression of SDC4 in mock or Chpf-overexpressed 4T1 cells. D. Silencing of SDC4 by siRNA suppresses CHPF-induced cell invasion. The mock or Chpf-overexpressed cells were transfected with non-targeting control siRNAs (Ctr si) or SDC4 specific siRNAs (SDC4 si) and subjected to transwell invasion assay. Data are expressed as the mean ± SD from three independent experiments. ***P < 0.001, *P < 0.05. The representative images are shown on the right. E. Silencing of SDC4 decreases CHPF-enhanced G-CSF accumulation. Protein levels of G-CSF and SDC4 were examined using western blotting. ACTB was used as loading the control. F. Relative mRNA expression of CHPF, SDC4, and G-CSF in the transfected cells. ***P < 0.001 compared to their control. G. CHPF enhanced G-CSF binding to SDC4. Immunoprecipitation (IP) of mock and Chpf-overexpressed 4T1 cell lysates with anti-SDC4 antibody or non-specific IgG control for 18 h at 4°C. Western blots of G-CSF and SDC4 after IP, and 40 μg of input proteins were shown at right. H. High expression of SDC4 with CHPF is associated with poor overall survival in breast cancer patients. Kaplan-Meier analysis of overall survival was performed according to gene expression. The median survival is shown on the right.

Article Snippet: G-CSF antibody (bs-1023R) was purchased from Bioss Inc.

Techniques: Immunoprecipitation, SDS Page, Staining, Western Blot, Expressing, Transfection, Transwell Invasion Assay, Binding Assay