Journal: Frontiers in Cell and Developmental Biology

Article Title: Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep

doi: 10.3389/fcell.2023.1177774

Figure Lengend Snippet: Top 20 DEPs derived from TMT proteome.

Article Snippet: For blocking, the anti-FCGR1A antibody (Bioss, Beijing, China) was incubated with semen (sperm concentration: 2–3 × 10 8 sperm/mL) at a working concentration of 50 μg/mL for 30 min.

Techniques: Derivative Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep

doi: 10.3389/fcell.2023.1177774

Figure Lengend Snippet: Correlation of FCGR1A with sperm fertilization ability. (A) Experimental design for the evaluation of the fertilization ability of FCGR1A-blocked sperm. (B) Difference between the sperm motility and viability after IVF with sperm obtained from different treatments. (C) Difference between the cleavage rate after IVF with sperm obtained from the different treatments. (D) Difference between blastocyst rates after in vitro fertilization with sperm from different treatments. FS represents fresh sperm after floating. FS + anti-FCGR1A represents fresh sperm blocked by anti-FCGR1A. PS represents programmed freezing sperm after floating. PS + anti-FCGR1A represents fresh sperm blocked by anti-FCGR1A. * p < 0.05, ** p < 0.001; ns, non-significance.

Article Snippet: For blocking, the anti-FCGR1A antibody (Bioss, Beijing, China) was incubated with semen (sperm concentration: 2–3 × 10 8 sperm/mL) at a working concentration of 50 μg/mL for 30 min.

Techniques: In Vitro

Journal: Frontiers in Cell and Developmental Biology

Article Title: Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep

doi: 10.3389/fcell.2023.1177774

Figure Lengend Snippet: List of possible correlations between the sperm motility, viability, and FCGR1A expression level.

Article Snippet: For blocking, the anti-FCGR1A antibody (Bioss, Beijing, China) was incubated with semen (sperm concentration: 2–3 × 10 8 sperm/mL) at a working concentration of 50 μg/mL for 30 min.

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep

doi: 10.3389/fcell.2023.1177774

Figure Lengend Snippet: Representative sperm mRNA and protein response mechanisms in the frozen semen of sheep. (A) Located in the sperm acrosomal and cytoplasm membrane, FCGR1A, ITGA3, and BET1 proteins are downregulated related to the pathway and function. (B) Located in the sperm nucleus, SLX4 is downregulated related to the pathway and function. (C) Schematic diagram showing the differences in sperm motility, viability, cleavage rate blocked, and unblocked sperm.

Article Snippet: For blocking, the anti-FCGR1A antibody (Bioss, Beijing, China) was incubated with semen (sperm concentration: 2–3 × 10 8 sperm/mL) at a working concentration of 50 μg/mL for 30 min.

Techniques: Membrane

Journal: EBioMedicine

Article Title: Rapid detection of bacterial infection using a novel single-tube, four-colour flow cytometric method: Comparison with PCT and CRP

doi: 10.1016/j.ebiom.2021.103724

Figure Lengend Snippet: A schematic diagram on how to calculate the FCBI-index value of the patient with microbiologically confirmed Streptococcus pneumoniae pneumonia using the novel FCBI-index method. (a) The incubation of the whole blood sample with the receptor-specific antibodies, red blood cell lysis, and flow cytometry run were carried out one after another in the same test tube. (b) During the flow cytometry run, the leukocyte population within gate R1 was first gated out from the side scatter (SSC)/MHCI bivariate histogram, after which gates R2 (red) and R3 (blue) were set around neutrophil and lymphocyte populations, respectively, in the SSC/CD64 bivariate histogram containing leukocytes inside gate R1. The stop count of 1000 monocytes inside the monocyte gate (black/R4) was used, leading to the collection of a total of 5000–20000 leukocytes. (c) For the purpose of data analysis, three additional bivariate histograms, namely SSC/CD35, SSC/CD64, and SSC/CD329 were made, showing mean fluorescence intensities (MFI, which correlate with the number of receptors on the cell surface) of gated neutrophil (red) and lymphocyte (blue) populations. In the SSC/CD35 histogram, B-lymphocytes expressing CD35 were left outside the lymphocyte gate. (d) The actual FCBI-index value was calculated by substituting the obtained receptor-specific MFI data from neutrophil and monocyte populations into the bacterial infection algorithm. A more detailed step-by-step description of the development of the FCBI-index method is presented in the method section and Supplementary Figs. 1–5. 1.54 × gCMF-HBSS: 1.54-fold calcium- and magnesium-free HBSS -buffer supplemented with 1.54 mg/mL gelatin.

Article Snippet: Fluorescence-conjugated receptor-specific monoclonal antibodies (test kit): Phycoerythrin (PE)-conjugated anti-human CD35 (Clone E11) was purchased from Ancell (ANC-184-050), PerCP-Vio700-conjugated anti-human CD64 (Clone 10.1.1) was purchased from Miltenyi Biotec (130-101-422), fluorescein isothiocyanate (FITC)-conjugated anti-human CD329 (Clone 191240) was purchased from R&D systems (FAB1139F) and AF647-conjugated anti-human MHC class I molecule (Clone W6/32) was purchased from R&D systems (FAB7098R). (Supplementary Table 5).

Techniques: Incubation, Red Blood Cell Lysis, Flow Cytometry, Fluorescence, Expressing, Infection

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD64 , REA987 , 50 , 130-116-195 , FITC , Miltenyi Biotec.

Techniques: Imaging

Journal: bioRxiv

Article Title: Neuronal C-Reactive Protein Mediates Neuropathic Pain by Activating Nociceptive FcγRI-Coupled Signaling

doi: 10.1101/2022.08.30.505953

Figure Lengend Snippet: (a) Western blot analysis of the protein levels of FcγRI and FcRγ (b) in DRGs from sham, CCI 7 d, CCI 15 d and 21 d rats after surgery. (b) Double immunofluorescence showing the cellular distribution of FcγRI (red) and the neuronal marker PGP9.5 (green) in DRGs from sham and CCI 21 d rats after surgery. (c) Double immunofluorescence showing the cellular distribution of FcRγ (red) with PGP9.5 (green) in DRGs from sham and 21 d rats after surgery. (d-e) Western blot analysis and quantification of the protein levels of pSyk in DRGs from sham, CCI 7 d and 15 d rats after surgery. (f) Double immunofluorescence showing the cellular distribution of pSyk (red) with the neuronal marker PGP9.5 (green) in DRGs from sham and CCI rats 15 d after surgery. (g) Proportion of pSyk-positive neurons in the DRGs of sham and CCI rats 15 d after surgery. * p < 0.05 versus sham, t test, n = 4/group. (h) Co-IP showed the interaction between FcγRI and Syk in DRG tissue from sham and CCI rats 15 d after surgery. (i) Co-IP showed the interaction between Syk and Vav1, pSrc and FcγRI in DRG tissue from sham and CCI rats 15 d after surgery. (j) Quantification of Co-IP of Syk, FcγRI, pSrc, and Vav1 in DRGs from sham and CCI rats 15 d after surgery. n = 6 rats/group, * p < 0.05 versus the sham group, one-way ANOVA. (k) Double immunofluorescence showing the cellular distribution of Vav1 in DRGs from sham and CCI rats 15 d after surgery. (l-m) Western blot analysis (l) and quantification (m) of the protein levels of pSrc and Src in DRGs from sham, CCI 7 d and 15 d rats after surgery. (n) Double immunofluorescence showing the cellular distribution of pSrc (red) with the neuronal marker PGP9.5 (green) in DRGs from sham and CCI rats 15 d after surgery. Scale bars: 50 μm in figure b, c, k and n; 30 μm in figure d and f, n = 4 rats/group. In figure a, d, e, i and m, n = 6 rats/group, * p < 0.05, ** p < 0.01 versus the sham group, one-way ANOVA.

Article Snippet: All membranes were blocked with 5% BSA in TBST for 1 hour at room temperature and subsequently incubated with rabbit anti-FcγRI (1:1000, Cat: 80016-R015, Sino Biological Inc.), rabbit anti-FcRγ (1:500, Cat: ab151986, Abcam), rabbit anti-Syk (1:1000, Cat: ab40781, Abcam), rabbit anti-pSyk (1:1000, Cat: PA5-36692, Thermo Fisher Scientific), rabbit anti-pSrc (1:1000, Cat: ab185617, Abcam), rabbit anti-Src (1:1000, Cat: ab109381, Abcam), rabbit anti-CRP (1:500, Cat: ab259862, Abcam), rabbit anti-Vav1 (1:500, Cat: D155205, Sangon Biotech), mouse anti-pNF-κB p65 (1:500, Cat: 13346, Cell Signaling Tech), rabbit anti-NLRP3 (1:1000, Cat: ab263899, Abcam), rabbit anti-IL-18 (1:1000, Cat: PAB16177, Abnova), rabbit anti-IL-1β (1:1000, Cat: ab254360, Abcam) or mouse anti-GAPDH (1:2000, Cat: ab8245, Abcam) primary antibodies.

Techniques: Western Blot, Immunofluorescence, Marker, Co-Immunoprecipitation Assay