Fig. 4: Mutant p53 promotes the immunosuppressive status of CD4⁺ T lymphocytes in the tumor microenvironment.

A Expression of endogenous p53^R273H in the exosomes isolated from HT-29 cell culture supernatant after pretreatment with serum starvation for 4 h and subsequent treatment with 10 ng/ml VEGF-A or 50 ng/ml bFGF for 12 h. B Expression of p53^R273H in the exosomes isolated from H1299 stable expressed p53^R273H cell culture supernatant after transfection with control, FLAG-K-RAS G12V, or H-RAS G12V plasmids. C Graphical quantification represents tumor growth rate in mice with time (n = 4) after an equal amount of 4T-1 control, Trp53^R270H-expressing cells were injected into the mammary gland of female BALB/c mice. D 4T-1 tumor-harboring mice were sacrificed on day 19. Representative images show tumor volume difference between Con and R270H. E Graphical quantification of the difference in tumor weight between Control and R270H mice (n = 4). F Flow cytometry analysis of CD45, CD3, and CD4 in T lymphocytes and the percentage of CD3⁺ CD4⁺ T lymphocytes isolated from R270H and its control mice. G Flow cytometry analysis of CD45, CD4, and PD-1 in T lymphocytes and the percentage of CD4⁺ PD1⁺ T lymphocytes isolated from R270H and its control mice. H Flow cytometry analysis of CD45, CD4, and TIGIT in T lymphocytes and the percentage of CD4⁺ TIGIT1⁺ T lymphocytes isolated from R270H and its control mice. Quantification of Interferon-γ⁺ (I) and tumor necrosis factor-α⁺ (J) cells as a percentage of CD4⁺ T cells, after 4 h of stimulation with PMA/Ionomycin, isolated from R270H and control mice.