Research Paper Volume 14, Issue 10 pp 4376—4389

Figure 4. has_circ_0069313 inhibited miR-325-3p induced foxp3 degradation. (A) The relative Foxp3 level in cells with indicated modifications (Student’s two-tailed paired test, ^***p < 0.001). (B) Immunoblot was applied to detect Foxp3 in cells with indicated modifications. (C) The detailed luciferase activity establishing strategy. (D) The luciferase activity reporting plasmid was transfected into HEK293T cells. The luciferase activity was measured and normalized (Student’s two-tailed paired test, ^***p < 0.001). (E) HEK293T was transfected with has_circ_0069313, has_circ_0069313 MUT, miR-325-3p mimic, miR-325-3p inhibitor alone or in combination. Luciferase activity was detected and normalized (Student’s two-tailed paired test, ^***p < 0.001). (F) SCC-9 cells were transfected with shRNA and shRNA plus miRNA inhibitor, relative Foxp3 level was detected and normalized (Student’s two-tailed paired test, ^***p < 0.001). (G) Cal-27 cells were transfected with OV plasmid, OV plasmid plus miRNA mimic, the relative Foxp3 and PDL1 level was detected and normalized (Student’s two-tailed paired test, ^***p < 0.001). (H) Immunoblot was applied to detect PDL1 and Foxp3 in cells with indicated modifications.