recombinant protein cd55 2009 cd  (R&D Systems)

Journal: Cell reports

Article Title: The expression profile and tumorigenic mechanisms of CD97 (ADGRE5) in glioblastoma render it a targetable vulnerability

doi: 10.1016/j.celrep.2023.113374

Figure Lengend Snippet: (A) Schematic of possible CD97 signaling mechanisms tested. Listed in red are the methods for testing. (B) Phosphoproteomic data collected from GBM samples identify five phosphorylation sites on the CD97 cytosolic C terminus. The heatmap depicts the percentage of GBM samples with the detected phosphorylation site. (C) Homogenous time-resolved fluorescence (HTRF) ratios from a β-arrestin recruitment assay performed after overexpression of WT CD97 or the ΔPS mutant. Two PDGCs (n = 2 for each) are compiled (n = 4 per condition; unpaired t test; ns p > 0.05; *p < 0.05; **p < 0.01). (D) Immunoblot for p-ERK1/ERK2 after overexpression of WT CD97 or the ΔPS mutant. (E) Bar graphs displaying densitometry ratios from (D) (n = 7; paired t test; ns p > 0.05; *p < 0.05). Two PDGCs (n = 3–4 for each) are compiled. (F) Bar graphs depicting the number of tumorspheres in a tumorsphere formation assay after CD97 knockdown followed by overexpression of shRNA-resistant forms of WT CD97 or the ΔPS mutant. The SCR shRNA groups used for normalization are not shown (n = 4 per PDGC; ANOVA F 5,12 = 80.79, p < 0.0001; Tukey’s multiple comparisons test: EV vs. WT: ***p < 0.001; EV vs. ΔPS: ns p > 0.05; WT vs. ΔPS: *p < 0.05). (G‒I) Single-cell RNA-/ATAC-seq data from showing expression of CD97 , CD55 , and THY1/CD90 . (J) A bar graph depicting the percentage of positive cells measured by flow cytometry after surface staining of PDGCs with fluorescein isothiocyanate (FITC)-conjugated antibodies against CD55, THY1/CD90, and an IgG control (n = 3 for each PDGC; two-way ANOVA F 2,20 = 95.11, p < 0.0001; Tukey’s multiple comparisons test: IgG vs. CD55: **p < 0.01; IgG vs. CD90: ****p < 0.00001; CD55 vs. CD90: ****p < 0.01). (K) Immunoblot for p-ERK1/ERK2 from CD97-overexpressing PDGCs plated on laminin or recombinant forms of putative ligands CD55 and THY1/CD90. (L) Quantification of densitometry ratios from immunoblots in (K) (n = 2–4 per PDGC; EV p-ERK/ERK: two-way ANOVA F 2,6 = 0.9450, p > 0.05; EV ERK/GAPDH: two-way ANOVA F 2,6 = 1.047, p > 0.05; CD97 overexpression p-ERK/ERK: two-way ANOVA F 2,16 = 12.48, p < 0.001; Tukey’s multiple comparisons test: laminin vs. CD55: ns, p < 0.05; laminin vs. CD90: ***p < 0.001; CD97 overexpression ERK/GAPDH: two-way ANOVA F 2,16 = 2.257, ns p > 0.05). Error bars indicate SEM.

Article Snippet: Wells were then additionally coated with recombinant human CD55 (Cat# 2009-CD-050, R&D) or recombinant human THY1/CD90 (Cat# 16897-HCCH, Sino Biological) at a concentration of 24 μg/mL overnight.

Techniques: Phospho-proteomics, Fluorescence, Over Expression, Mutagenesis, Western Blot, Tube Formation Assay, Knockdown, shRNA, Expressing, Flow Cytometry, Staining, Control, Recombinant

Journal: Cell reports

Article Title: The expression profile and tumorigenic mechanisms of CD97 (ADGRE5) in glioblastoma render it a targetable vulnerability

doi: 10.1016/j.celrep.2023.113374

Figure Lengend Snippet:

Article Snippet: Wells were then additionally coated with recombinant human CD55 (Cat# 2009-CD-050, R&D) or recombinant human THY1/CD90 (Cat# 16897-HCCH, Sino Biological) at a concentration of 24 μg/mL overnight.

Techniques: Recombinant, Modification, Disruption, Activation Assay, Knock-Out, shRNA