Journal: International Journal of Molecular Sciences

Article Title: Exosomal Carboxypeptidase E (CPE) and CPE-shRNA-Loaded Exosomes Regulate Metastatic Phenotype of Tumor Cells

doi: 10.3390/ijms23063113

Figure Lengend Snippet: Characterization of exosomes loaded with CPE-shRNA. ( A ) Schematic showing the strategy of loading and transfer of CPE-shRNA via exosomes. Exosomes were isolated from supernatant media of HEK293T cells (ExoHEK) infected with adenovirus encoding either CPE-shRNA or CTRL-shRNA, fused to GFP. HCC97H cells treated with these modified exosomes exhibited green fluorescence, validating the transfer of CPE-shRNA through the exosomes. Representative images showing GFP fluorescence in target HCC97H cells, treated with either ExoHEK-CPE-shRNA or ExoHEK-CTRL-shRNA are included. Scale bar = 100 µm. ( B ) Graph showing the concentration and size distribution of ExoHEK-CPE-shRNA and ExoHEK-CTRL-shRNA, as determined by NanoSight analysis.

Article Snippet: For experiments targeting HCC97H with CPE-shRNA-loaded exosomes, HEK293T cells were infected with adenovirus carrying either CPE-shRNA-GFP or control-shRNA-GFP (Vector Biolabs, Cat# shADV-229236, Malvern, PA, USA) at MOI 25 for 48–72 h. After 5–6 h of infection, the culture media were replaced to remove viral particles present in the infection media.

Techniques: shRNA, Isolation, Infection, Modification, Fluorescence, Concentration Assay

Journal: MethodsX

Article Title: Development and evaluation of mouse anti-Ara h 1 and Ara h 3 IgE monoclonal antibodies for advancing peanut allergy research

doi: 10.1016/j.mex.2023.102470

Figure Lengend Snippet: Dose dependency of each IgE monoclonal antibody against CPE in a sandwich ELISA.

Article Snippet: All plates were screened for anti-CPE IgE antibody-producing hybridomas by ELISA kits (Cat# 3063, Chondrex, Inc. WA, USA), according to the manufacturer's instructions.

Techniques: Sandwich ELISA

Journal: MethodsX

Article Title: Development and evaluation of mouse anti-Ara h 1 and Ara h 3 IgE monoclonal antibodies for advancing peanut allergy research

doi: 10.1016/j.mex.2023.102470

Figure Lengend Snippet: Dose dependency of each IgE monoclonal antibody against CPE in an indirect ELISA.

Article Snippet: All plates were screened for anti-CPE IgE antibody-producing hybridomas by ELISA kits (Cat# 3063, Chondrex, Inc. WA, USA), according to the manufacturer's instructions.

Techniques: Indirect ELISA

Journal: MethodsX

Article Title: Development and evaluation of mouse anti-Ara h 1 and Ara h 3 IgE monoclonal antibodies for advancing peanut allergy research

doi: 10.1016/j.mex.2023.102470

Figure Lengend Snippet: Summary of the Application of Anti-CPE IgE Monoclonal Antibodies.

Article Snippet: All plates were screened for anti-CPE IgE antibody-producing hybridomas by ELISA kits (Cat# 3063, Chondrex, Inc. WA, USA), according to the manufacturer's instructions.

Techniques: Indirect ELISA, Sandwich ELISA

Journal: The EMBO journal

Article Title: Antitumor T-cell function requires CPEB4-mediated adaptation to chronic endoplasmic reticulum stress.

doi: 10.15252/embj.2022111494

Figure Lengend Snippet: Figure 2. CPEB4 is required for T-cell effector function but not for early activation, proliferation, or induction of differentiation.

Article Snippet: Membranes were blocked in 5% milk and incubated with antibodies against CPEB4 (Mouse Monoclonal ERE149C, 1:500, homemade), b-Actin-HRP (1:15,000, ab49900, Abcam), vinculin (1:1,000, ab130007 Abcam), DDIT4 (1:1,000, 10,638-1-AP, Proteintech), HERPUD2 (1:500, sc398583, Santa Cruz), phospho-eiF2a (s51) (1:1,000, ab32157, Abcam), total eiF2a (1:1,000, 9722S, Cell Signaling), CHOP (1:1,000, CST5554T, Cell Signaling), and BiP (1:1,000, C50B12, Cell Signaling).

Techniques: Activation Assay

Journal: The EMBO journal

Article Title: Antitumor T-cell function requires CPEB4-mediated adaptation to chronic endoplasmic reticulum stress.

doi: 10.15252/embj.2022111494

Figure Lengend Snippet: Figure 3. CPEB4 protein is upregulated in tumor-infiltrated effector CD8 T cells and upon T-cell activation ex vivo.

Article Snippet: Membranes were blocked in 5% milk and incubated with antibodies against CPEB4 (Mouse Monoclonal ERE149C, 1:500, homemade), b-Actin-HRP (1:15,000, ab49900, Abcam), vinculin (1:1,000, ab130007 Abcam), DDIT4 (1:1,000, 10,638-1-AP, Proteintech), HERPUD2 (1:500, sc398583, Santa Cruz), phospho-eiF2a (s51) (1:1,000, ab32157, Abcam), total eiF2a (1:1,000, 9722S, Cell Signaling), CHOP (1:1,000, CST5554T, Cell Signaling), and BiP (1:1,000, C50B12, Cell Signaling).

Techniques: Activation Assay, Ex Vivo