Figure 3.

Pdia3 deficiency intrinsically impairs Th1 and Th17 program

(A) Representative pictures of the thymus, mesenteric lymph nodes (mLN), and spleen from 8- to 12-week-old WT and KO mice. (B–F) Splenic Tn cells isolated from WT and KO mice aged 10 weeks were activated with plate-coated anti-CD3/CD28 (10 μg/mL) antibodies for 72 h. (B and C) The percentage of proliferated CD4 T cells was defined by Ki67 staining (B) and CFSE assay (C). (D) Apoptosis of T cells was determined by Annexin V/PI staining. (E and F) Expression of IFN-γ (E) and IL-17A (F) was examined in activated CD4 T cells. (G and H) Tn cells purified from WT and KO mice were cultured under Th1- or Th17-polarized conditions in vitro. Flow cytometry analysis of Th1 (G) and Th17 (H) cell frequencies. (I) The scheme for the co-culture experiments. (J) The percentage of proliferated CD4 T cells was defined by CFSE or cell-trace violet (CTV) staining. (K and L) The percentage of IFN-γ-producing Th1 cells (K) and IL-17A-producing Th17 cells (L) after co-culturing. The experiments were repeated at least three times.