Fig. 5

ADQ inhibits TAM/CXCL1-induced differentiation and immunosuppressive function of Tregs in vitro. a CCK-8 assay indicated that 20–200 μg/ml ADQ treatment for 24 h exhibited a moderate inhibitory effect on naive CD4⁺ T cell proliferation. b The differentiation changes of naive CD4⁺ T cells when treated as indicated for 24 h were detected by flow cytometry. C–d 293 T cells (c) and naive CD4⁺ T cells (d) were treated as indicated for 24 h. Western blot (c) and immunofluorescence assays (d) were conducted to detect the FOXP3 expression levels in 293 T cells and naive CD4⁺ T cells. Scale bar = 3 μm. e–f Flow cytometry assay indicated that pretreatment of TAMs with 20 μg/ml ADQ for 24 h partially abrogated TAM/CXCL1-induced immunosuppressive function of Tregs on CD8⁺ T cells. The proliferation activities, apoptosis (e), as well as granzyme B and perforin secretion levels (f) of CD8⁺T cells were detected by flow cytometry. The LDH activity of the 4T1 cells (f) was investigated by the colorimetric assay. N = 3. *p < 0.05; **p < 0.01