Journal: Frontiers in Immunology

Article Title: Targeting Pin1 to overcome immunosuppressive tumor microenvironment in MSS colorectal cancer

doi: 10.3389/fimmu.2025.1677029

Figure Lengend Snippet: Pin1 promotes Tregs chemotactic migration through the CCL3-CCR5 axis. (A) Correlation analysis of Pin1 with CCL chemokines expression. (B, C) qRT-PCR and ELISA assessing CCL3 expression in SW480 and HT29 cells after Pin1 knockdown. (D) Schematic representation of PBMCs isolated from human peripheral blood; Treg cells sorted by flow cytometry, these populations co-cultured with siNC- or siPin1-CRC cells, respectively. (E, F) Level of Treg markers (CD25 + CD127 - ) on PBMCs in the co-culture system detected by flow cytometry. (G) Effect of supernatants from Pin1-knockdown cells treated with recombinant CCL3 protein on Treg chemotactic capacity. (H) Correlation between CCL3 and CCR5 expression in CRC tissues. (I) CCR5-neutralizing antibodies abolish differences in Treg chemotaxis between the siNC and siPin1 groups.

Article Snippet: The concentrations of CCL3 in culture media were evaluated using Human CCL3 (MIP-1a) ELISA Kit (liankebio, cat.no.

Techniques: Migration, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Knockdown, Isolation, Flow Cytometry, Cell Culture, Co-Culture Assay, Recombinant, Chemotaxis Assay

Journal: Frontiers in Immunology

Article Title: Targeting Pin1 to overcome immunosuppressive tumor microenvironment in MSS colorectal cancer

doi: 10.3389/fimmu.2025.1677029

Figure Lengend Snippet: Pin1 regulates CCL3 expression to promote Treg recruitment and CAFs activation through the NF-κB signaling pathway. (A) Validation of Pin1 interaction with p65 using CO-IP. (B) Co-localization of Pin1 with P65 confirmed using IF. (C) The main components of the NF-κB signaling pathway on knockdown of Pin1, as determined using WB. (D) Representative images of p65 IHC staining in subcutaneous tumors in mice (N.S. + IgG, Pin1 + IgG) (n=5 per group). (E) qRT-PCR assessed CCL3 expression in SW620 cells after Pin1 overexpression with/without NF-κB inhibitor treatment. (F) NF-κB inhibitor eliminated Pin1-overexpression-induced differences in Treg chemotaxis. (G) NF-κB inhibitor abrogated the activating effect of Pin1 overexpression on CAFs, as analyzed via IF.

Article Snippet: The concentrations of CCL3 in culture media were evaluated using Human CCL3 (MIP-1a) ELISA Kit (liankebio, cat.no.

Techniques: Expressing, Activation Assay, Biomarker Discovery, Co-Immunoprecipitation Assay, Knockdown, Immunohistochemistry, Quantitative RT-PCR, Over Expression, Chemotaxis Assay