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. Author manuscript; available in PMC: 2015 Jan 8.
Published in final edited form as: J Immunol Methods. 2011 Sep 6;376(0):1–12. doi: 10.1016/j.jim.2011.08.023

Figure 3. Expression and phosphorylation of PKC-θ in Tregs and non-Tregs following in vitro stimulation with anti-CD3ε mAb, SEB, and OVA.

Figure 3

Using flow cytometry, the intracellular expression and phosphorylation of PKC-θ was assessed in Tregs vs. non-Tregs. The stimuli anti-CD3ε mAb (column A), SEB (column B), and OVA-peptide (column C) were added in different doses (0, 0.01, 0.1, or 1 µg/ml) to cultures of splenocytes harvested from WT or OT-II transgenic mice. Following incubation times from 30 min to 2 hours, the expression and phosphorylation of the intracellular signaling molecule PKC-θ was analyzed. The charts represent the quantitative data of the relative fluorescence intensity (RFI) for stained intracellular PKC-θ and p-PKC-θ in Tregs and non-Tregs. Data represent one of two independent experiments. N=3 individual mice per group. Mean ± SEM given, #p<0.05 1 µg/ml vs. 0 µg/ml stimulation at same time point, *p<0.05 vs. 30 min stimulation at 1µg/ml.