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. 2022 Jun 28;14(13):3164. doi: 10.3390/cancers14133164

Figure 5.

Figure 5

Ectopic GPR56 expression inhibits migration of primary healthy T cells. (A) Primary healthy T cells ectopically expressing GPR56 were generated by transducing T cells with lentivirus containing pRRL-SFFV-GPR56-iGFP or pRRL-SFFV-iGFP. After 3 days the GPR56 expression was evaluated by flow cytometry. (B) Migration in response to chemoattractant CXCL12a was performed with primary healthy T cells lentivirally transduced with pRRL-SFFV-GPR56-iGFP or an empty vector. Primary healthy T cells were loaded in a volume of 100 µL on transwell filters with a pore size of 5 µm. SDF-1 (CXCL12) was added as chemoattractant to the lower compartment at a concentration of 0–500 ng/mL in a total volume of 200 µL of migration buffer. After 2.5 h incubation at 37 °C, cells in the lower compartment were harvested and quantified by flow cytometry. The percentage of migration was calculated considering the migration of the primary healthy T cells transduced with the empty vector as 100%. A two-tailed unpaired t test was used to compare the levels of migration.