Journal: Cell Reports Methods

Article Title: Discovery of antibodies targeting multipass transmembrane proteins using a suspension cell-based evolutionary approach

doi: 10.1016/j.crmeth.2023.100429

Figure Lengend Snippet: Key resources table

Article Snippet: The following staining antibodies were used: (i) APC-conjugated mouse anti-human PD-L1 antibody (BioLegend, clone MIH2, 1:50 dilution), (ii) Alexa647-conjugated mouse anti-human CXCR2 antibody (BioLegend clone 5E8/CXCR2, 1:20 dilution), (iii) Alexa647-conjugated mouse anti-human GLP-1R antibody (R&D Systems clone 197920, 1:20 dilution), (iv) Alexa647-conjugated mouse anti-human GCGR antibody (R&D Systems clone 591929, 1:20 dilution), (v) APC-conjugated mouse anti-human CXCR4 antibody (Invitrogen clone 12G5, 1:20 dilution), (vi) APC-conjugated mouse anti-human CCR5 antibody (Invitrogen clone NP-6G4, 1:20 dilution).

Techniques: Virus, Recombinant, Expressing, Modification, Infection, Transfection, Affinity Chromatography, Transduction, Plasmid Preparation, Sequencing, Software

Journal: Cell reports

Article Title: Insulin Secretion Depends on Intra-islet Glucagon Signaling.

doi: 10.1016/j.celrep.2018.10.018

Figure Lengend Snippet: Figure 1. Exogenous Glucagon Infusions Stimulate Insulin Secretion through the Glucagon and the GLP-1 Receptor in Perfused Mouse Pancreata (A, B, E, and F) Insulin secretion from perfused pancreata during infusion of glucagon (0.1–10 nM) at 12 mM glucose in control mice with or without Ex9 (A, n = 8), Glp1r/ mice (B, n = 6), Gcgrbcell/ mice (E, n = 5), and global Gcgr/ mice with or without Ex9 (F, n = 8). (C, D, G, and H) Mean insulin output averaged over 10 min before and during addition of glucagon to perfused pancreata of control mice (C), Glp1r–/– (D), Gcgrbcell-/- (G), and Gcgr-/- (H) mice. Significance was tested by 1-way ANOVA (repeated measures) comparing stimulated output versus respective basal output. Data are represented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. See also Figure S1.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, Peptides, and Recombinant Proteins Glucagon Sigma-Aldrich G2044 Exendin 9-39 Bachem 4017799 L-arginine Sigma-Aldrich A6969 Experimental Models: Cell Lines COS-7 cells ATCC, Virginia, USA CRL-1651 Experimental Models: Organisms/Strains Mouse: DT-Gcg Pedersen et al., 2013 n/a Mouse: Glp-1r / Scrocchi et al., 1996 n/a Mouse: Gcgrbcell / Daniel Drucker, University of Toronto n/a Mouse: Gcgr / Gelling et al., 2003 n/a Recombinant DNA Mouse Gcgr DNA Origene MC203290 mouse Glp1r DNA Origene MC216256 Other In-house ELISA for active GLP-1 Vilsbøll et al., 2003 n/a Glucagon antiserum for glucagon radioimmunoassay Orskov et al., 1991 4305 Insulin antiserum for insulin radioimmunoassay Brand et al., 1995 2006-3

Techniques: Control

Journal: Endocrine

Article Title: GLP-1 receptor agonists show no detrimental effect on sperm quality in mouse models and cell lines

doi: 10.1007/s12020-025-04245-4

Figure Lengend Snippet: GLP-1R agonists have no direct effect on sperm motility and the testicular cell proliferation. A : Protein expression of GLP-1R was assessed in four testicular cell lines and testicular tissue by western blot. Mouse β-cell line Min6 cells were used as positive control. B : Immunohistochemical staining of GLP-1R in testicular tissue. Scale bar = 50 μm. The cells in the dashed box are enlarged on the right. C : Gene expression data of GLP-1R in human pancreatic islet, testis, testis germ cell, testis intersitial, Leydig cell. The data were generated by using BioGPS. D–L : Sperm suspensions collected from cauda epididymis of male C57BL/6 J mice were treated with liraglutide (Lira, 100 nmol/L), semaglutide (Sema, 100 nmol/L), or saline (as control [Ctrl]). n = 8 per group. Sperm motility was assessed. D : Rapid progressive motility (grade A sperm). E : Progressive motility (grade A + B sperm). F : Total motility (grade A + B + C sperm). G : Velocity along the straight‑line path. H : Velocity along the curvilinear path. I : Velocity along the average path. J : Beat cross frequency. K : Straightness. L : Amplitude lateral head displacement. M : Mouse spermatogonial cell line GC-1 cells, spermatocyte cell line GC-2 cells, Leydig cell line TM3 cells, and Sertoli cell line TM4 cells were treated with different concentrations of liraglutide or semaglutide (0–100 nmol/L) for 72 h. Cell proliferation was analyzed using CCK-8 kit ( n = 4 per group). All data are presented as mean ± SD. Statistical analysis was conducted by one-way ANOVA in D–L, or by two-way ANOVA in M

Article Snippet: The membranes were probed with the following primary antibodies overnight at 4 °C: Goat polyclonal anti-GLP-1R antibody (1:1000; OriGene) and mouse monoclonal anti-β-actin antibody (1:10,000; C1313, Applygen, Beijing, China).

Techniques: Expressing, Western Blot, Positive Control, Immunohistochemical staining, Staining, Gene Expression, Generated, Saline, Control, CCK-8 Assay

Journal: Endocrine

Article Title: GLP-1 receptor agonists show no detrimental effect on sperm quality in mouse models and cell lines

doi: 10.1007/s12020-025-04245-4

Figure Lengend Snippet: GLP-1R ablation has no effect on testicular morphology and sperm motility. The metabolic indicators and sperm parameters in eight-week-old male Glp1r −/− mice and the littermate wildtype (WT) mice ( n = 8 per group). A : Body weight. B : Random blood glucose level. C : Blood glucose during oral glucose tolerance test (OGTT). D : HE staining of the testicular tissue. Scale bar = 50 μm. The cells in the dashed box are enlarged on the right. E–M : The sperm count and motility assessment. E : Sperm concentration. F : Rapid progressive motility (grade A sperm). G : Progressive motility (grade A + B sperm). H : Velocity along the straight‑line path. I : Velocity along the curvilinear path. J : Velocity along the average path. K : Beat cross frequency. L : Straightness. M : Amplitude lateral head displacement. All data are presented as mean ± SD. Statistical analysis was conducted by Student’s t- test in A–C and E–M, ** P < 0.01, *** P < 0.001

Article Snippet: The membranes were probed with the following primary antibodies overnight at 4 °C: Goat polyclonal anti-GLP-1R antibody (1:1000; OriGene) and mouse monoclonal anti-β-actin antibody (1:10,000; C1313, Applygen, Beijing, China).

Techniques: Staining, Concentration Assay