Human C5L2/GPR77 Antibody (650918)

Detection of C5L2/GPR77 in HEK293 Human Cell Line Transfected with Human C5L2/GPR77 and eGFP by Flow Cytometry.

HEK293 human embryonic kidney cell line transfected with (A) C5L2/GPR77 or (B) irrelevant protein, and eGFP were stained with Mouse Anti-Human C5L2/GPR77 Monoclonal Antibody (Catalog # MAB10254) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). Quadrant markers were set based Mouse IgG2B Isotype Control Antibody staining (Catalog # MAB0041, data not shown). View our protocol for Staining Membrane-associated Proteins.

Detection of C5L2/GPR77 by Flow Cytometry

Characterization and functional analysis of CAFs in CRC. (A) Flow cytometric analysis illustrating the identification of fibroblasts isolated from samples from CRC patients using the markers CD10 and GPR77, highlighting the heterogeneity within the CAF population. The experiment was conducted in triplicate to ensure the reliability and reproducibility of the results. (B) Morphological examination of CAFs through light microscopy, depicting their elongated spindle or stellate shapes with thin cytoplasmic extensions and prominent actin stress fibers. The adjacent panels show the immunofluorescent staining results for alpha -SMA and vimentin with a comparison of the expression levels in CAFs and NFs, demonstrating significantly higher expression in CAFs; magnification, ×400. (C) Gene expression microarray data showcasing significant changes in the expression profiles of the proteins, including chemokines and cytokine receptors, secreted by CAFs compared with NFs, with alterations marked as greater-than-five-fold increases. (D) Additional microarray results focusing on distinct clusters of upregulated genes related to tumor invasion and metastasis in CAFs. (E) qRT-PCR results displaying elevated mRNA levels of vimentin, fibroblast activation protein, alpha -SMA, SDF-1, and CXCR4 in CAFs extracted from CRC tissues; these were corroborated by corresponding increases in protein levels, reinforcing the pro-tumorigenic role of these markers in the tumor microenvironment. This experiment was conducted in triplicate to ensure the reliability and reproducibility of the results. (Scale bar: 50 μm). Data are shown as representative images and numerical data are represented as the mean ± SD of each group of cells from three separate experiments. *** p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39195225), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of C5L2/GPR77 by Flow Cytometry

Characterization of CAFs and NFs obtained from clinical surgical tissues from patients with head and neck cancer (HNC) (a) Morphological comparisons between CAFs and NFs from a representative HNC case showed that CAFs (right panel) consisted of more cytoplasmic protrusions than NFs (left panel). Photographs were captured at 40× magnification. (b) Quantitative PCR (left panel) of the culture medium showed a significantly higher expression of vimentin and alpha -SMA in CAFs than in NFs. Western blot analysis (right panel) also demonstrated that levels of vimentin and alpha -SMA were significantly higher in CAFs than in NFs. (c) Flow cytometric analysis of the cell surface markers, CD10 and GPR77, showed a marked increase in their expression in CAFs compared to NFs. (d) A heat map of the gene microarray of NFs and CAFs showed that there were several differences in the expression profile of the secreted genes. The arrow indicates a marked discrepancy of the relative mRNA levels of CCL11 in CAF compared with NF. (e) The RT-PCR (left panel) and ELISA (right panel) analysis showed an increased expression of CCL11 in CAFs compared with that in NFs. (f) Western blot analysis showed that the protein level of CCL11 was higher in CAFs than in NFs in cell lysates. (g) Western blot analysis showed a higher CCL11 expression in CAF-CM compared to NF-CM. The asterisk indicated a significant difference (*: p < 0.05; **: p < 0.01) between experimental and control groups. Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/2072-6694/14/13/3141), licensed under a CC-BY license. Not internally tested by R&D Systems.