Journal: JCI Insight

Article Title: Inhibition of cell surface GRP78 and activated α 2M interaction attenuates kidney fibrosis

doi: 10.1172/jci.insight.183998

Figure Lengend Snippet: ( A ) Schematic outline of the study. Created in BioRender. Trink, J. (2025) https://BioRender.com/2rod2yk ( B ) Fα2M significantly reduced fibrosis measured as trichrome, PSR, and fibronectin, as well as the myofibroblast marker α–smooth muscle actin (α-SMA). ( C ) Treatment with Fα2M also prevented Smad3 and FAK activation (measured by phosphorylation at S473/475 and Y397, respectively), as well as increased YAP expression compared with control IgG1 ( n = 7–9; * P < 0.05, ** P < 0.01, **** P < 0.0001; # P < 0.05 significant by t test; scale bar represents 20 μm).

Article Snippet: Three separate studies were conducted: (a) treatment with C38 (made in-house) or the isotype control IgG2b (BioXcell, BE0086) antibodies; (b) Fα2M (made in-house) or the isotype control IgG1 (BioXcell, BE0083) antibodies; or (c) P19 or scrambled control peptide (both made in-house).

Techniques: Marker, Activation Assay, Phospho-proteomics, Expressing, Control

Journal: Cell Reports

Article Title: Flow Cytometry of Mouse and Human Adipocytes for the Analysis of Browning and Cellular Heterogeneity

doi: 10.1016/j.celrep.2018.08.006

Figure Lengend Snippet:

Article Snippet: The human/mouse Anti-UCP1 antibody (MAB6158, monoclonal Mouse IgG 2B Clone # 536435, R&D Systems) was conjugated to Alexa647 (ThermoFisher Alexa Fluor 647 antibody labeling kit A20186) and incubated with adipocytes at 1:300 for 1 h. Mouse IgG2B Alexa Fluor 647-conjugated Isotype Control antibody (R&D systems) was used as a control.

Techniques: Control, Recombinant, Blocking Assay, Antibody Labeling, Software, Flow Cytometry

Journal: International journal of molecular sciences

Article Title: Effect of Flavonoids on MCP-1 Expression in Human Coronary Artery Endothelial Cells and Impact on MCP-1-Dependent Migration of Human Monocytes.

doi: 10.3390/ijms242216047

Figure Lengend Snippet: Figure 7. Effect of recombinant MCP-1 (A) and a neutralizing antibody (nab) directed against MCP-1 (B) on monocyte migration. Migration of THP-1 cells was determined with Boyden chamber assay toward increasing MCP-1 concentrations (A) or toward CM from HCAEC (B). In (A), medium containing recombinant MCP-1 added to the lower chamber was used to induce monocyte migration. In (B), CM from HCAEC previously treated with IL-1β or vehicle for 24 h was incubated with MCP-1 antibody (1 µg/mL) or the appropriate isotype control IgG2B (1 µg/mL) for 1 h after removal from the cells. Thereafter, migration of THP-1 monocytes toward CM was initiated. In both settings (A,B), migration was run for 6 h. Monocyte migration toward vehicle-containing medium (A) or toward CM of IL-1β-treated HCAEC containing IgG2B (B) were set to 100%. Data are presented as means ± SEM of n = 5–6 (A) or n = 6 (B) of three independent experiments each. ** p ≤0.01, *** p ≤0.001 vs. vehicle control; one-way ANOVA with Dunnett’s post hoc test (A). *** p ≤0.001 vs. vehicle control (leftmost white bar); ### p ≤0.001 vs. IL-1β-stimulated cells; one-way ANOVA with Bonferroni’s post hoc test (B).

Article Snippet: IgG2B Isotype Control (#MAB004; RRID:AB_357346) and CCL2/JE/MCP-1 Antibody (#MAB679; RRID:AB_2071559) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: Recombinant, Migration, Boyden Chamber Assay, Incubation, Control

Journal: Translational Oncology

Article Title: HER2 as a potential therapeutic target on quiescent prostate cancer cells

doi: 10.1016/j.tranon.2023.101642

Figure Lengend Snippet: Impact of T-DM1 treatment on metastasis free survival in a prostate cancer left ventricle injection xenograft model. (A) Experimental design for the in vivo experiment. GFP/luciferase-expressing PC3 cells were injected into male SCID mice by left ventricle intracardiac (i.c.) injection. Human IgG (15 mg/kg) or T-DM1 (15 mg/kg) were injected by intraperitoneal (i.p.) injection every 3 days until 12 days after tumor injection IgG: n = 12. T-DM1: n = 14. (B) Kaplan-Meier analysis of time to formation of metastases visible by bioluminescence imaging or death. (C) Representative bioluminescence images of control mice (human IgG injected) and T-DM1 treated mice 124 days after tumor injection. (D) Antibody dependent cellular cytotoxicity (ADCC) assays using flow cytometry. PC3 cells were labeled with DiD for subsequent identification, and were treated with T-DM1 before addition of splenocytes from SCID mice. PC3 cells were identified as DiD positive and non-viable cells were identified as positive for DAPI. (E) Dead PC3 cells (%) quantified from data in panel (D) Data represent mean ± S.D. (N=3).

Article Snippet: Monoclonal mouse IgG2B (R&D systems, Cat #: IC0041V) was used as isotype control.

Techniques: Injection, In Vivo, Luciferase, Expressing, Imaging, Control, Flow Cytometry, Labeling