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. 2019 Apr 12;10:753. doi: 10.3389/fimmu.2019.00753

Figure 4.

Figure 4

APDS variants enhance p85α-iOPN interactions. (A) Immunoprecipitation of WT or Y524S p110δ with p85α. 293T cells were co-transfected with HA-p110δ and Myc-p85α, and p85α was immunoprecipitated with an anti-Myc affinity gel. Results are representative of two experiments. (B) Co-immunoprecipitation of p85α with iOPN in the presence of WT or Y524S p110δ. 293T cells were co-transfected with WT or variant HA-p110δ, iOPN-FLAG and the indicated amount of Myc-p85α. iOPN was immunoprecipitated with an anti-FLAG affinity gel. Results are representative of two experiments. (C) Co-immunoprecipitation of iOPN with p85α from control and patient CD4+ T cell blasts. Results are representative of two experiments. (D) Intracellular OPN expression in nuclear and cytoplasmic fractions of transduced (GFP+) cells. CD4+ T cells from a healthy donor ere transduced with WT or Y524S p110δ. Blotting for GAPDH shows proper fractionation. Non-specific low molecular weight bands from the OPN blot show equal protein loading in the nuclear fractions. Results are representative of two experiments. (E) Model showing that enhanced interaction of iOPN with p85α in the presence of the variant p110δ drives TFH differentiation.