Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Upregulation of p16 INK4A in Peripheral White Blood Cells as a Novel Screening Marker for Colorectal Carcinoma

doi: 10.31557/APJCP.2022.23.11.3753

Figure Lengend Snippet: (a) Immunofluorescence staining of CD3+ peripheral immune cells showing DAPI staining (i), p16 INK4A staining (ii), CD3 staining (iii) and merged image (iv), (b) Immunofluorescence staining of CD14+ peripheral immune cells showing DAPI staining (i), p16 INK4A staining (ii), CD3 staining (iii) and merged image (iv), (c) Scatter plot of p16 INK4A MFI of healthy controls and p16 INK4A MFI of CRC group in CD3+ and CD14+ cells. The bar on the scatter plot represents mean value

Article Snippet: Anti-CD3 primary mouse antibody (Cell signaling) and anti-CD14 primary mouse antibody (Cell signaling) were diluted 1:500.

Techniques: Immunofluorescence, Staining

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Upregulation of p16 INK4A in Peripheral White Blood Cells as a Novel Screening Marker for Colorectal Carcinoma

doi: 10.31557/APJCP.2022.23.11.3753

Figure Lengend Snippet: (a) Comparison of p16INK4A positive cells in white blood cells, CD3+ cells and CD14+ cells between CRC group and healthy control group, (b) Correlation of p16 INK4A -positive cells percentage and p16 INK4A mean fluorescent intensity of CRC patients and healthy controls in white blood cells, (c) Correlation of p16 INK4A -positive cells percentage and p16 INK4A mean fluorescent intensity of CRC patients and healthy controls in CD3+ cells, (d) Correlation of p16 INK4A -positive cells percentage and p16 INK4A mean fluorescent intensity of CRC patients and healthy controls in CD14+ cells

Article Snippet: Anti-CD3 primary mouse antibody (Cell signaling) and anti-CD14 primary mouse antibody (Cell signaling) were diluted 1:500.

Techniques: Comparison, Control

Journal: Cell Death & Disease

Article Title: M1 macrophage-derived exosomes and their key molecule lncRNA HOTTIP suppress head and neck squamous cell carcinoma progression by upregulating the TLR5/NF-κB pathway

doi: 10.1038/s41419-022-04640-z

Figure Lengend Snippet: A The infiltration levels of immune cells in HNSCC. B Prediction of overall survival according to high- and low-infiltration levels of M0, M1 and M2 macrophages. THP-1-derived M0, M1 and M2 macrophages were confirmed by flow cytometry using ( C ) CD14 plus CD86 and ( D ) CD14 plus CD163 ( n = 3). E The exosomes were extracted from three types of macrophages and confirmed by electronic microscopy and Western blot analysis using CD9 and CD63. F RT-qPCR was conducted to test the expression of HOTTIP in M0, M1 and M2 macrophages ( n = 3). RT-qPCR was conducted to test the expression of HOTTIP in wild type M1 macrophages, HOTTIP-overexpressed and HOTTIP-knockdown M1 macrophages ( G ) and their exosomes ( H ) ( n = 3). Data are presented as mean ± SD. Results were analyzed by One-way ANOVA with a post hoc t -test. Significance: * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The cells were dissociated at 4 °C with PBS-EDTA, resuspended in calcium- and magnesium-free PBS and incubated at 4 °C with an anti-CD14 primary antibody (human: 11-0149-42, eBioscience, USA; mice: 123307, Biolegend, American) for 1 h. After the cells were washed with PBS, they were incubated with a fluorescein-conjugated anti-mouse IgG secondary antibody at 4 °C for 45 min. Then, the cells were suspended in a PBS solution.

Techniques: Derivative Assay, Flow Cytometry, Microscopy, Western Blot, Quantitative RT-PCR, Expressing, Knockdown

Journal: Cell Death & Disease

Article Title: M1 macrophage-derived exosomes and their key molecule lncRNA HOTTIP suppress head and neck squamous cell carcinoma progression by upregulating the TLR5/NF-κB pathway

doi: 10.1038/s41419-022-04640-z

Figure Lengend Snippet: A–D Flow cytometry assay detected CD14 + CD86 + M1 and CD14 + CD163 + M2 phenotype in circulating blood of nude mice bearing with HOTTIP-knockdown Hep-2 cells ( n = 3). HOTTIP-knockdown Hep-2 cells suppressed the polarization of M1 monocytes ( A , C ) but induced the polarization of M2 ( B , D ). E – H Flow cytometry assay detected M1 and M2 phenotypes in the blood of tumor-bearing nude mice treated by M1 exosomes ( n = 3). M1 exosomes induced the polarization of M1 monocytes ( E , G ) but inhibited the polarization of M2 ( F , H ). Data are presented as mean ± SD. Results were analyzed by One-way ANOVA with a post hoc test. Significance: ns not significant, ** P < 0.01, *** P < 0.001.

Article Snippet: The cells were dissociated at 4 °C with PBS-EDTA, resuspended in calcium- and magnesium-free PBS and incubated at 4 °C with an anti-CD14 primary antibody (human: 11-0149-42, eBioscience, USA; mice: 123307, Biolegend, American) for 1 h. After the cells were washed with PBS, they were incubated with a fluorescein-conjugated anti-mouse IgG secondary antibody at 4 °C for 45 min. Then, the cells were suspended in a PBS solution.

Techniques: Flow Cytometry, Knockdown

Journal: Cell systems

Article Title: Gut-Liver physiomimetics reveal paradoxical modulation of IBD-related inflammation by short-chain fatty acids

doi: 10.1016/j.cels.2020.02.008

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Transwell membranes containing epithelial monolayers on the apical side and macrophages/DCs on the basolateral side were washed with FACS buffer (PBS with 2%FBS) and stained with primary mouse anti-human CD14 Clone MφP9 (BD Biosciences, cat. no. 347490).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Cell Isolation, Software, Luminex