Figure 4. ZFP91 deficiency promotes T cell activation and proliferation.

(A) Volcano plot comparing global gene expression profiles between Zfp91^+/+ Cd4-Cre (WT) and Zfp91^fl/fl Cd4-Cre (KO) CD90.2⁺ T cells stimulated with anti-CD3 and anti-CD28 antibodies for 24 hours. The red dots represented the transcripts with increasing expression values, whereas the blue dots signify transcripts with decreasing expression values in Zfp91^fl/fl Cd4-Cre CD90.2⁺ T cells. (B) Gene ontology enrichment analysis of upregulated and downregulated gene sets in Zfp91^fl/fl Cd4-Cre (KO) T cells stimulated with anti-CD3 and anti-CD28 antibodies for 24 hours. (C) Heatmap of upregulated genes associated with T cell activation and proliferation in ZFP91-deficient T cells relative to gene expression in WT T cells. (D) ELISA of IFN-γ and IL-2 in supernatants of Zfp91^+/+ Cd4-Cre and Zfp91^fl/fl Cd4-Cre CD8⁺ T cells stimulated with anti-CD3 and anti-CD28 antibodies (n = 4). (E) Flow cytometric analysis of Ki-67 expression in Zfp91^+/+ Cd4-Cre and Zfp91^fl/fl Cd4-Cre CD4⁺ or CD8⁺ T cells stimulated with anti-CD3 and anti-CD28 antibodies for 48 hours (n = 3). (F) Zfp91^+/+ Cd4-Cre and Zfp91^fl/fl Cd4-Cre CD4⁺ or CD8⁺ T cells (3 × 10⁵ cells per well) were labeled with CFSE and cultured with irradiated splenocytes depleted of T cells (1 × 10⁵ cells per well) in the presence of anti-CD3 and anti-CD28 antibodies for 72 hours for T cell proliferation assays (n = 3). Experiments were independently repeated 3 times. Data are presented as the mean ± SEM. *P < 0.05 and **P < 0.01, by 2-tailed Student’s t test.