Fig 2: The FKBP4/NR3C1/TMEM173 signaling pathway involved in DC abundance(A) Distribution of FKBP4, NR3C1, and TMEM173 expression across immune subtypes in pan-cancer analysis using the TISIDB database. (B) Distribution of FKBP4, NR3C1, and TMEM173 expression across immune subtypes only in BC using the TISIDB database. (C) Spearman correlations between expression of FKBP4, NR3C1, or TMEM173 and infiltration level of DCs in BC using the TIMER database. (D) Spearman correlations between expression of FKBP4, NR3C1, or TMEM173 and abundance of DCs in BC using the TISIDB database.

Fig 3: GR correlates with PD-L1 expression, low MHC-I expression, and poor survival in human PDAC.a Representative immunohistochemical (IHC) staining of GR in the normal pancreatic duct and PDAC. Scale bars, 100 µm (left), 50 µm (middle), and 50 µm (right). b Quantification of GR-positive and GR-negative cases of PDAC and adjacent normal pancreatic tissue. n = 101 patients. Statistical significance was determined by a two-tailed Fisher’s exact test. c GR (encoded by NR3C1) mRNA levels in paired normal pancreatic tissue and PDAC based on the GSE15471 dataset. n = 36 patients. Statistical significance was determined by a two-tailed paired t-test. d, e Representative IHC staining (d) and statistical analysis (e) of the correlation of GR protein levels with PD-L1, MHC-I, and CD8 proteins levels in patients with PDAC. n = 101 patients. f, g Correlation of NR3C1 (encoding GR) mRNA levels with CD274 (encoding PD-L1; f) and HLA-A (g) mRNA levels in PDAC based on TCGA data. n = 178 patients. h Kaplan–Meier curves of overall survival of pancreatic cancer patients stratified by GR protein levels based on IHC. n = 101 patients. i Kaplan–Meier curves of overall survival of pancreatic patients stratified by GR (encoded by NR3C1) mRNA levels. Data were obtained from the ICGC (International Cancer Genome Consortium). n = 95 patients. Statistical significance was determined by the log-rank test in h and i. j Plasma cortisol levels in healthy volunteers (n = 122) and PDAC patients (n = 82). Statistical significance was determined by a two-tailed unpaired t-test. Error bars are s.e.m. k–m Correlation of plasma cortisol levels with PD-L1 (k), MHC-I (l), or GR (m) protein levels in pancreatic tumors. n = 32 patients. Statistical significance was determined by a two-tailed Pearson correlation test in e–g and k–m. Source data are provided as a Source Data file.

Fig 4: Modulation of PD-L1 and MHC-I by GR is a common regulatory mechanism in PDAC cells.a Immunoblotting of GR, MHC-I, and GAPDH in human PDAC cell lines. b, c qPCR analysis of PD-L1, HLA-A, HLA-B, and HLA-C in HPAC (b) and BXPC-3 (c) cells with or without MIFE (20 µM, 72 h) treatment. n = 3 technical replicates. d, e Immunoblotting of PD-L1, MHC-I, and GAPDH in HPAC (d) and BXPC-3 (e) cells with or without MIFE treatment with the indicated doses for 72 h. f Left panel: qPCR analysis of Nr3c1, Pd-l1, H-2k, H-2d, and B2m in GR-knockdown HY24409 cells. Right panel: immunoblotting of GR and Gapdh in GR-knockdown HY24409 cells. g qPCR analysis of Pd-l1, H-2k, H-2d, and B2m in HY24409 cells with or without MIFE (20 µM, 48 h) treatment. h qPCR analysis of Pd-l1, H-2k, H-2d, and B2m in HY24409 cells treated with DEX (100 nM, 4 h) and MIFE (100 nM, 4 h), alone or in combination. n = 3 technical replicates in f–h. i–k Representative flow cytometry plots and quantification of cell-surface PD-L1 (i), MHC-I (j), and B2M (k) in HY24409 cells with or without MIFE (20 µM, 48 h) treatment. n = 3 biological replicates. l Left panel: qPCR analysis of Pd-l1, H-2k, H-2d, B2m, and Nr3c1 in GR-knockdown HY19636 cells. n = 3 technical replicates. Right panel: immunoblotting of GR and Gapdh in GR-knockdown HY19636 cells. m–o Representative flow cytometry plots and quantification of cell-surface PD-L1 (m), MHC-I (n), and B2M (o) in GR-knockdown HY19636 cells. n = 3 biological replicates. p qPCR analysis of Pd-l1, H-2k, H-2d, and B2m in HY19636 cells with or without MIFE (20 µM, 48 h) treatment. n = 3 technical replicates. q–s Representative flow cytometry plots and quantification of cell-surface PD-L1 (q), MHC-I (r), and B2M (s) in HY19636 cells with or without MIFE (20 µM, 48 h) treatment. n = 3 biological replicates. Statistical significance in b, c, and f–s was determined by a two-tailed unpaired t-test. Error bars are s.e.m. Source data are provided as a Source Data file.

Fig 5: FKBP4 binds to NR3C1 and FKBP4 inhibition triggers nuclear translocation of NR3C1. (A–D) co-IP and western blot assays using antibodies as indicated for binding between endogenous FKBP4 and NR3C1 in T47D and BT549 cells. (E–H) T47D and BT549 cells were transfected with siRNAs or plasmids. Cells were harvested, and the nuclear and cytoplasmic fractions were separated. Proteins were analyzed by immunoblotting using anti-NR3C1, anti-FKBP4, anti- GAPDH, or anti-histone antibody. (I, J) T47D and BT549 cells were transfected with FKBP4 siRNAs. Immunofluorescent staining was carried out using anti-NR3C1 and anti-FKBP4. DAPI staining was performed to show the nuclei (n = 3 independent biological replicates). The arrowheads show the NR3C1 nuclear localization under FKBP4 inhibition. Scale bar = 20 µm.