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. 2014 Oct 30;6(5):1290–1299. doi: 10.4161/mabs.29758

Figure 6.

Figure 6.

Effects of PD-1 engagement on phospho PTEN and casein kinase 2 levels in CD4+ effector memory T cells stimulated by anti-CD3 and CD28SA. Human CD4+ TEMs were stimulated with anti-CD3 mAb (CD3, 5 μg/ml); NIB1412 (NIB1412, 10 μg/ml); rPD-L1 (rPD-L1, 10μg/ml); with anti-CD3 mAb and rPD-L1 (CD3 + rPD-L1) or with NIB1412 and rPD-L1 (NIB1412 + rPD-L1). At the end of 72 h, the cells were lysed and extracted protein separated by SDS-PAGE. pPTEN – S380/T382/383 (A) and CK2 (B) protein levels were assessed by immunoblotting and β-actin was used as loading control. All blots are representative of four independent experiments. pPTEN and CK2 levels were quantified by densitometry, normalized to β-actin and expressed as a percentage of pPTEN or CK2 levels in untreated cells. Data are represented as mean ± SEM of four independent experiments. Statistical analysis was performed using unpaired t test (*p < 0.05, **p < 0.01). Phospho PTEN – pPTEN; Casein Kinase 2 – CK2