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. 2017 Mar 27;37(2):BSR20160485. doi: 10.1042/BSR20160485

Figure 2. PKCζ regulates T-cell functional phenotype potential during maturation of human CB T-cells in culture.

Figure 2

Human CB T-cells were maturated to CD45RO+/RA– phenotype by culturing with addition of PHA (day 1) and IL-2 (day 3) over 7 days. (A) PKCζ knockdown was achieved by nucleofection of CB T-cells with specific shRNA. Nucleofection with PKCζ shRNA led to the loss of PKCζ in T-cells, as demonstrated by Western blot of three separate CB samples. (B) Evaluating day 8 of cultures for the level of T-cell maturation by the levels of expression of CD45RA/CD45RO. (C,D) T-cells developed on PKCζ-deficient background and then stimulated with PHA/PMA were examined for (C) proliferation and (D) cytokine (TNFα, IFNγ and IL-13) production. The effects of PKCζ deficiency on cytokine synthesis are expressed as % of control production where the control values were 460 ± 115.5 pg/ml, 350.5 ± 125 pg/ml and 96 ± 57 pg/ml respectively (D). Results are expressed as mean ± S.E.M. of three experiments. Statistics: *, P<0.05; **, P<0.01.