Figure 5.

PDIA3 is upregulated in CD4 T cells via the TCR/Wnt5a-Ca²⁺-NFAT1 axis

(A) The putative binding site in the Pdia3 promoter region for NFAT1 was predicted with a relative profile score threshold of 80%. (B) ChIP-PCR analysis of the NFAT1 binding activity to the Pdia3 promoter. (C) Luciferase reporter assays for the Pdia3 promoter conducted in HEK 293T cells. (D and E) Tn cells from WT and KO mice polarized to Th1(D) and Th17(E) cells in the presence of recombinant Wnt5a. (F) The expression of NFAT1 and PDIA3 was measured by immunoblotting in the presence of recombinant Wnt5a at the indicated concentrations. (G) The scheme to validate the upstream pathway of PDIA3. (H) The expression of NFAT1 and PDIA3 detected by Western blot analysis in the presence of recombinant Wnt5a and/or iNFAT1. (I–L) Tn cells from WT mice were skewed to Th1 (I and J) and Th17 (K and L) in the presence of recombinant Wnt5a and/or iNFAT1. (M and N) Flow cytometry analysis of calcium ions by detecting Fluo-4 (M) and Rhod-2 (N). (O) Western blot results of NFAT1 and PDIA3 expression in the presence of recombinant Wnt5a and/or EDTA. (P) The protein levels of NFAT1 and PDIA3 in the presence of recombinant Wnt5a and/or MSAB.