Journal: Biochimica et Biophysica Acta. Molecular Basis of Disease

Article Title: Single cell RNA-seq resolution revealed CCR1 + /SELL + /XAF + CD14 monocytes mediated vascular endothelial cell injuries in Kawasaki disease and COVID-19

doi: 10.1016/j.bbadis.2023.166707

Figure Lengend Snippet: The differential activation of CD14 and CD16 monocytes responding to endothelial and epithelial dysfunction. The profile scores of molecules mediating the interplays of classic monocytes with vascular endothelial cells (A-B) and non-classic monocytes with alveolar epithelial cells (C—D). E. Immunostaining for CDH5 and γH2AX in coronary artery cryo-section between KD patient and healthy donor, indicating endothelial injuries in KD. F. Expression level of CCR1 , DYSF , SELL , LMNB1, and XAF1 in CD14 classical monocytes among five groups. G. UMAP projection of CCR1 , DYSF , SELL , LMNB1, and XAF1 positive cells, respectively . H. UMAP projection of CCR1 , SELL and XAF1 triple positive monocytes among five groups. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, Wilcoxon rank-sum test, adjusted for Bonferroni post hoc test, had been applied.

Article Snippet: Cell smears fixated in 4 % PFA were blocked in blocking buffer (PBS containing 1 % BSA, 0.1 % Triton X-100, 5 % goat serum) for 1 h at room temperature, followed by incubation with rabbit anti-Dysferlin (1:200, Abcam, cat # ab124684), rabbit anti-XAF1 (1:200, Abcam cat #ab2254) antibodies and mouse anti-CD14 (1:200, Abcam cat #ab181470) diluted in PBS containing 1 % BSA, 5 % goat serum at 4 °C overnight.

Techniques: Activation Assay, Immunostaining, Expressing

Journal: Biochimica et Biophysica Acta. Molecular Basis of Disease

Article Title: Single cell RNA-seq resolution revealed CCR1 + /SELL + /XAF + CD14 monocytes mediated vascular endothelial cell injuries in Kawasaki disease and COVID-19

doi: 10.1016/j.bbadis.2023.166707

Figure Lengend Snippet: SELL+/CCR1+/XAF1+ CD14 monocytes enhanced the adhesion and damages to endothelial cells. A. Flow cytometry identified the higher percentages and median fluorescence intensity of SELL, CCR1, and LMNB1 in KD and COV. B. Immunostaining for CD14 and DYSF, XAF1 in isolated PBMCs. C. The ratio of DYSF positive CD14 monocytes in total classic monocytes among KD, COV, FLU, and healthy donors. And the median fluorescence intensity of XAF1 in CD14 monocytes among KD, FLU, and healthy donors. D-E. THP-1 had been stained with Calcein AM and transfected siRNAs of CCR1 , DYSF , SELL , LMNB1 , and XAF1 before co-culture with HUVECs. Then the numbers of adhesion THP-1 with HUVECs were counted by every 20× field view. F. The expressions of TNFa and IL6 in HUVECs, and the ratio of γH2AX + cells in HUVECs after co-cultured with THP-1, which had been transfected with siRNAs of CCR1 , DYSF , SELL , LMNB1 , and XAF1 . G. The inhibition of SELL , CCR1 and XAF1 together in THP-1 significantly reduced the adhesion between monocytes and endothelial cells. H. Collaborated inhibiton of SELL , CCR1 and XAF1 in THP-1 decreased the expression of IL6 and TNF-α in HUVECs with lower ratio of rH2AX+ cells after co-culture with THP-1. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, Two-way analysis of variance with Bonferroni post hoc test was performed to analyze data. Bar, 100 μm.

Article Snippet: Cell smears fixated in 4 % PFA were blocked in blocking buffer (PBS containing 1 % BSA, 0.1 % Triton X-100, 5 % goat serum) for 1 h at room temperature, followed by incubation with rabbit anti-Dysferlin (1:200, Abcam, cat # ab124684), rabbit anti-XAF1 (1:200, Abcam cat #ab2254) antibodies and mouse anti-CD14 (1:200, Abcam cat #ab181470) diluted in PBS containing 1 % BSA, 5 % goat serum at 4 °C overnight.

Techniques: Flow Cytometry, Fluorescence, Immunostaining, Isolation, Staining, Transfection, Co-Culture Assay, Cell Culture, Inhibition, Expressing

Journal: Frontiers in Physiology

Article Title: Using Imaging Mass Cytometry to Define Cell Identities and Interactions in Human Tissues

doi: 10.3389/fphys.2021.817181

Figure Lengend Snippet: List of antibodies used on human kidney tissue.

Article Snippet: CD14 , Monocytes/Macrophages , Rabbit , EPR3653 , 144Nd , 1:50 , Fluidigm , .

Techniques: Membrane

Journal: Poultry Science

Article Title: Research note: Turkey ( Meleagris gallopavo ) semen CD14 + macrophages – management, key features, and classification – a pilot study with optimized protocols ☆

doi: 10.1016/j.psj.2025.105397

Figure Lengend Snippet: The visualization of turkey macrophages using immunofluorescence (IF) staining. A. A representative turkey semen CD14 + type 1 macrophages (CD14 + iNOS + , akin CD14 + Mф1); B. A representative turkey semen CD14 + type 2 macrophages (CD14 + IL10 + , akin CD14 + Mф2); C. A representative small mononucleated semen CD14 + Mф with an absence of vacuoles; d-F. Hypothetical process of multinucleated giant cell formation from two mononucleated small semen CD14 + Mф; G. Multinucleated giant semen CD14 + Mф with internalized sperm cells. Full arrows indicate vacuoles, and empty arrows indicate selected sperm. Images were captured using a confocal Laser Scanning Microscope (LSM800; Carl-Zeiss, Germany) equipped with an Airyscan super-resolution module (40 × /1.2 NA W c-apochromat) supported by Zen Blue Pro 2.6 software (Carl-Zeiss; Germany).

Article Snippet: Next, cells were triple-washed with PBS and covered with primary mouse anti-CD14 antibodies (orb434888, Biorbyt, United Kingdom, 1:50 in PBS).

Techniques: Immunofluorescence, Staining, Laser-Scanning Microscopy, Software

Journal: Poultry Science

Article Title: Research note: Turkey ( Meleagris gallopavo ) semen CD14 + macrophages – management, key features, and classification – a pilot study with optimized protocols ☆

doi: 10.1016/j.psj.2025.105397

Figure Lengend Snippet: A. Turkey semen CD14 + Mф measured by flow cytometry I-III: The representative cytogrammes show the gating strategy for identifying the different CD14 + Mф subpopulations. B. I-X: Consecutive photographs (10-min intervals) of turkey semen Mф phagocyting zymosan particle imaged using fluorescence microscopy with an Axio Observer. Z1/7 fluorescence microscope (Carl Zeiss, Inc., Oberkochen, Germany) equipped with ZEN 2.3 blue edition software (Carl Zeiss). C. Turkey semen Mф 24 h after seeding (magnification 200 ×), and D. after culture washing (magnification 400 ×). The high incidence of sperm in the cell pool decreases when cultured in high-glucose DMEM and after cell washing. Full arrows indicate selected Mф and empty arrows indicate sperm.

Article Snippet: Next, cells were triple-washed with PBS and covered with primary mouse anti-CD14 antibodies (orb434888, Biorbyt, United Kingdom, 1:50 in PBS).

Techniques: Flow Cytometry, Fluorescence, Microscopy, Software, Cell Culture