Journal: Frontiers in Immunology
Article Title: Defective Localization With Impaired Tumor Cytotoxicity Contributes to the Immune Escape of NK Cells in Pancreatic Cancer Patients
doi: 10.3389/fimmu.2019.00496
Figure Lengend Snippet: Chemokines and their receptor expression in PDAC patients. (A) qRT-PCR analysis of multiple chemokine expression in tumor tissues and non-tumor tissues harvested from patients undergoing resection surgery. (B) Flow cytometry analysis of the expression of chemokine receptors in circulating NK cells from patients vs. that from healthy donors. (C) CXCR2 expression on NK cells in healthy donor was gated on CD56hi and CD56lo populations (left), and their percentages in HD and PDAC patients are summarized as a table (right), with the average level of CXCR2 expression on CD56hi and CD56lo NK cells shown as bar graphs (D) Flow cytometry analysis of CXCR2 expression on NK cells from PBMC and TIL of patients with PDAC. (E) Schematic diagrams of lentivirus construction expressing CXCR2 gene; CXCR2 gene was cloned upstream of RSV-copGFP cassette in the transfer vector pCDH-521A. The level of CXCR2 expression on transduced NK cells was shown by FACs. (F) Migration assay was performed using vector-or CXCR2-transduced patients' NK cells targeting MIA PaCa-2 (left) or Patients' tumor cells (right). Bar graph shows the percentage of NK cells migrated toward tumor targets. Data shown here are representative of three independent experiments. Statistical significance was determined by one-way ANOVA ( * p ≤ 0.05; *** p < 0.001).
Article Snippet: Full-length human CXCR2 was obtained from Addgene (cat #66260, MA) and amplified by polymerase chain reaction. pCDH-521A lentiviral plasmids carrying CXCR2 were co-transfected with packaging plasmids into 293 FT cells using CaCl 2 , and supernatant was collected after 72 h of culture.
Techniques: Expressing, Quantitative RT-PCR, Flow Cytometry, Clone Assay, Plasmid Preparation, Migration
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